Novel tetragalnac and peptide containing conjugates and methods for delivery of oligonucleotides

ABSTRACT

Disclosed herein is a modular composition comprising 1) an oligonucleotide; 2) one or more tetraGalNAc ligands of Formula (I), which may be the same or different; optionally, 3) one or more linkers, which may be the same or different; 4) one or more peptides independently selected from Table  3 , which may be the same or different; and optionally, 5) one or more targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. patent application Ser. No. 15/481,942, filed on Apr. 7, 2017, which is continuation application of U.S. patent application Ser. No. 14/398,369, filed on Oct. 31, 2014, which is a national-stage application of PCT Application No. PCT/US2013/039072, filed on May 1, 2013, which claims the benefit of priority to U.S. Provisional Patent Application Ser. No. 61/641,741, filed May 2, 2012; all of which are incorporated by reference herein in their entirety.

BACKGROUND OF THE INVENTION

Scientific efforts focused on the delivery of oligonucleotides systemically for therapeutic purposes are ongoing. Three highlighted approaches to oligonucleotide delivery include 1) lipid nanoparticle (LNP) encapsulation, 2) polymer conjugation and 3) single chemical conjugation. Single chemical conjugation typically employs a targeting ligand or a lipid or a solubilizing group or an endosomolytic peptide or a cell penetrating peptide and/or a combination of two or all four attached to an oligonucleotide. Linkers may be present in the conjugate as well as other functionalities. Single chemical conjugates are known and attachment of the oligonucleotide occurs either at the 5′- or 3′-end of the oligonucleotide, at both ends, or internally. See WO2005/041859, WO2008/036825, and WO2009/126933.

Considerable amount of literature evidence supports the hypothesis that the major hurdles for oligonucleotide delivery are cell uptake and endosomal escape. There remains a need for additional single chemical conjugates that can provide effective delivery efficiency, cell uptake and/or endosomal escape.

SUMMARY OF THE INVENTION

Single chemical conjugates comprising tetraGalNAc and peptides disclosed herein have surprising properties of effective delivery efficiency, cell uptake and/or endosomal escape.

In one embodiment, a modular composition disclosed herein comprises: 1) a single stranded or double stranded oligonucleotide; 2) one or more tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different:

wherein X is —O—, —S—, —CR¹R²— or —NR¹—, wherein R¹ and R² are each independently selected from the group consisting of hydrogen and C1-C6alkyl; n is 1, 2, 3, or 4; and the bond with “

” indicates point of attachment; optionally, 3) one or more linkers, which may be the same or different; 4) one or more peptides independently selected from Table 3, which may be the same or different; and optionally, 5) one or more targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents. In one embodiment, R¹ and R² are each independently selected from the group consisting of hydrogen, methyl and ethyl. In another embodiment, R¹ and R² are each hydrogen.

In one embodiment, the tetraGalNAc ligand has Formula (II) wherein X, R¹, R² and n are as defined above. In another embodiment, the tetraGalNAc ligand has Formula (III) wherein X, R¹, R² and n are as defined above:

In another embodiment, a modular composition comprises: 1) a single stranded or double stranded oligonucleotide; 2) 1-8 tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different, wherein X is —O—, —S—, —CH₂— or —NH—; and n is 1, 2, 3, or 4; 3) 1-24 linkers, which may be the same or different; 4) 1-8 peptides independently selected from Table 3, which may be the same or different; and optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents. the tetraGalNAc ligand has Formula (II) wherein X, R¹, R² and n are as defined above.

In another embodiment, a modular composition comprises: 1) a single stranded or double stranded siRNA; 2) 1-8 tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different, wherein X is —O—, —S—, —CH₂— or —NH—; and n is 1, 2, 3, or 4; 3) 1-24 linkers, which may be the same or different; 4) 1-8 peptides independently selected from Table 3, which may be the same or different; and optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.

In another subset of the above embodiments, the linkers are attached to the oligonucleotide or siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the oligonucleotide or siRNA.

In another subset of the above embodiments, the tetraGalNAc ligands and/or the peptides are attached to the oligonucleotide or siRNA optionally via linkers.

In another subset of the above embodiments, the tetraGalNAc ligands and/or the peptides are attached to the oligonucleotide or siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the oligonucleotide or siRNA; and the tetraGalNAc ligands and/or the peptides are attached to the oligonucleotide or siRNA optionally via linkers.

In another subset of the above embodiments, X of Formula (I), (II) or (III) is —O—, —S—, or —CH₂—; and n is 1, 2 or 3.

In another subset of the above embodiments, X of Formula (I), (II) or (III) is —O— or —CH₂— and n is 1 or 2.

In another subset of the above embodiments, X of Formula (I), (II) or (III) is —O— and n is 1 or 2.

In another subset of the above embodiments, X of Formula (I), (II) or (III) is —CH₂— and n is 1 or 2.

In another subset of the above embodiments, the composition comprises 1-6 tetraGalNAc ligands, or more specifically, 1-4 tetraGalNAc ligands, which may be the same or different.

In another subset of the above embodiments, the composition comprises 1-6, peptides, or more specifically, 1-4 peptides, which may be the same or different.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded; and the tetraGalNAc ligands are attached to the guide strand or the passenger strand of the oligonucleotide or siRNA at different 2′-positions of the ribose rings.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded; and the tetraGalNAc ligands are attached to the guide strand or the passenger strand of the oligonucleotide or siRNA at different terminal 3′ and/or 5′-positions.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded; and the tetraGalNAc ligands are attached to both the guide strand and the passenger strand of the oligonucleotide or siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded; and the peptides are attached to the guide strand or the passenger strand of the oligonucleotide or siRNA at different 2′-positions of the ribose rings of the siRNA.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded; and the peptides are attached to the guide strand or the passenger strand of the oligonucleotide or siRNA at different terminal 3′ and/or 5′-positions.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded; and the peptides are attached to both the guide strand and the passenger strand of the oligonucleotide or siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions.

In another subset of the above embodiments, the tetraGalNAc ligands and the peptides are attached to the same strand of the oligonucleotide or siRNA.

In another subset of the above embodiments, the tetraGalNAc ligands and the peptides are attached to different strands of the oligonucleotide or siRNA.

In another subset of the above embodiments, the tetraGalNAc ligands and the peptides are attached to the same or different strands of the oligonucleotide or siRNA via linkers.

In another subset of the above embodiments, each linker is independently selected from Table 1.

In another subset of the above embodiments, each linker is independently selected from Table 2.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded; and the optional targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents are attached to the same or different strands of the oligonucleotide or siRNA.

In one embodiment, a modular composition comprises 1) a double stranded siRNA; 2) 1-8 tetraGalNAc ligands of Formula (IV), (V) or (VI), which may be the same or different:

3) 1-24 linkers independently selected from Table 1, which may be the same or different; 4) 1-8 peptides independently selected from Table 3, which may be the same or different; and, optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.

In another embodiment, a modular composition comprises 1) a double stranded siRNA; 2) 1-4 tetraGalNAc ligands of Formula (IV), (V) or (VI), which may be the same or different; 3) 1-12 linkers independently selected from Table 1, which may be the same or different; 4) 1-4 peptides independently selected from Table 3, which may be the same or different; and, optionally, 5) 1-4 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents; wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA optionally via linkers.

In one subset of the above embodiments, the tetraGalNAc ligands and the peptides are attached to the same strand of the siRNA via linkers.

In another subset of the above embodiments, the tetraGalNAc ligands and the peptides are attached to different strands of the siRNA via linkers.

In one embodiment, a modular composition comprises 1) a double stranded siRNA; 2) 1-4 tetraGalNAc ligands of Formula (IV), (V) or (VI), which may be the same or different; 3) 1-12 linkers independently selected from Table 2, which may be the same or different; 4) 1-4 peptides independently selected from Table 4, which may be the same or different; and, optionally, 5) 1-4 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents; wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA via linkers.

In one subset of the above embodiment, the tetraGalNAc ligands and the peptides are attached to the same strand of the siRNA via linkers.

In one subset of the above embodiment, the tetraGalNAc ligands and the peptides are attached to different strands of the siRNA via linkers.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Non-limiting examples of modular compositions comprising double stranded oligonucleotides with terminal conjugations.

FIG. 2. Non-limiting examples of modular compositions comprising double stranded oligonucleotides with terminal conjugations.

FIGS. 3A-3B. Non-limiting examples of modular compositions comprising double stranded oligonucleotides with internal and/or terminal conjugations are shown in FIG. 3A to FIG. 3B.

FIG. 4. Generic structures of each nucleotide [O_(n)] or [O_(n′)] that contain a linker (L-P and/or L-G).

FIGS. 5A-1-5D. Scheme 2 as shown in FIG. 5A-1 to FIG. 5D for preparing B Conjugates (Ex. 3-6).

FIGS. 6A-6B. Scheme 3 as shown as FIG. 6A to FIG. 6B for preparing Conjugates B6-P32 and B8-seq32 (Ex. 7-8). The figures disclose SEQ ID NO: 32.

FIGS. 7A-7I. Scheme 4 as shown in FIG. 7A, FIG. 7B and FIG. 7C for preparing B9, B10-seq32 and B11-seq32. The figures disclose SEQ ID NO: 32.

Scheme 5 as shown in FIG. 7D-1 and FIG. 7D-2, FIG. 7E and FIG. 7F for preparing B-13-seq13-b compound. The figures disclose SEQ ID NO: 13.

Scheme 6 as shown in FIG. 7G-1 to FIG. 7G-2 for preparing B16-seq32 and B17-seq32-b compound. FIG. 7H-1, FIG. 7H-2, and FIG. 7I show the preparation of B15-seq32 and B16-seq32-b. FIGS. 7H-1 to 7I disclose SEQ ID NO: 32.

FIGS. 8A-8D. Scheme 7 as shown in FIG. 8A to FIG. 8D for preparing C1 to C3, C4-seq32 and C6-seq32 compound. The figures disclose SEQ ID NO: 32.

FIGS. 9A-9E. Scheme 8 as shown in FIG. 9A to FIG. 9E for preparing C7 to C10, C11-seq32 and C12-seq32 compound. The figures disclose SEQ ID NO: 32.

FIGS. 10A-10D. Scheme 9 shown in FIG. 10 A to FIG. 10D for preparing C13, C14-seq32 and C15-seq32-a compound. The figures disclose SEQ ID NO: 32.

FIGS. 11A-11D. Scheme 10 as shown in FIG. 11A to FIG. 11D for preparing D1, D3 and D4.

FIGS. 12A-1-12B-2. Scheme 11 as shown in FIG. 12A-1 to FIG. 12B-2 for preparing D5-seq32 and D7-seq32 compound. The figures disclose SEQ ID NO: 32.

FIGS. 13A-13H-2. Scheme 12 as shown in FIG. 13A to FIG. 13H-2 for preparing E compounds.

FIGS. 14A-1-14B-2. Scheme 13 as shown in FIG. 14A-1 to FIG. 14B-2 for preparing E8-seq 137 and E10-seq137e compounds. The figures disclose SEQ ID NO: 137.

FIGS. 15A-15E-2. Scheme 14 as shown in FIG. 15A to FIG. 15E-2 for preparing F compounds. The figures disclose SEQ ID NO: 463.

FIGS. 16A-1-16B-2. Scheme 15 as shown in FIG. 16A-1 to FIG. 16B-2 for preparing F6seq 463-f compound. The figures disclose SEQ ID NO: 463.

FIGS. 17A-1-17D-2. Scheme 16 as shown in FIG. 17A-1 to FIG. 17D-2 for preparing G compounds. The figures disclose SEQ ID NO: 489.

FIGS. 18A-1-18B-2. Scheme 17 as shown in FIG. 18A-1 to FIG. 18B-2 for preparing G compounds. The figures disclose SEQ ID NO: 489.

FIGS. 19A-19I-2. Scheme 19 as shown in FIG. 19A to FIG. 19I-2 for preparing H10-seq32-h compound. The figures disclose SEQ ID NO: 32.

FIGS. 20A-1-20E-2. Scheme 20 as shown in FIG. 20A-1 to FIG. 20E-2 for preparing I10-seq1681-f compound. The figures disclose SEQ ID NOS 1737, 1737-1739, 1737, 1737, and 1737, respectively, in order of appearance.

FIGS. 21A-21H-2. Scheme 21 as shown in FIG. 21A to FIG. 21H-2 for preparing J9-seq26-i compound. The figures disclose SEQ ID NO: 26.

FIGS. 22A-1-22D-2. Scheme 22 as shown in FIG. 22A-1 to FIG. 22D-2 for preparing K6 seq 74-b compound. The figures disclose SEQ ID NO: 74.

FIGS. 23A-23C-2. Scheme 23 as shown in FIG. 23A to FIG. 23C-2 for preparing L11-seq 463-j compound. The figures disclose SEQ ID NO: 463.

FIGS. 24A-1-24B-2. Scheme 24 as shown in FIG. 24A-1 to FIG. 24B-2 for preparing M4-seq-j compound. The figures disclose SEQ ID NO: 463.

FIGS. 25A-25B-2. Scheme 25 as shown in FIG. 25A to FIG. 25B-2 for preparing N4-seq 283-k compound. The figures disclose SEQ ID NO: 283.

FIGS. 26A-1-26B-2. Scheme 26 as shown in FIG. 26A-1 to FIG. 26B-2 for preparing O3-seq 463-k compound. The figures disclose SEQ ID NO: 463.

FIGS. 27A-1-27B-2. Scheme 27 as shown in FIG. 27A-1 to FIG. 27B-2 for preparing P2-seq-32-k compound. The figures disclose SEQ ID NO: 13.

FIGS. 28-1-28-2. Scheme 28 as shown in FIG. 28-1 to FIG. 28-2 for preparing P2-seq 32-m compound. The figures disclose SEQ ID NO: 74.

FIGS. 29A-1-29C-2. Scheme 29 as shown in FIG. 29A-1 to FIG. 29C-2 used to prepare Q3-seq74-b compound. The figures disclose SEQ ID NO: 74.

FIGS. 30A-30E-3. Scheme 30 as shown in FIG. 30A to FIG. 30E-3 for preparing R4-seq 27-I compound. The figures disclose SEQ ID NO: 27.

FIGS. 31A-31B. Scheme 32 as shown in FIG. 31A and FIG. 31B for preparing tetraGalNAc-siRNA conjugates.

FIGS. 32A-32B. Scheme 33 as shown in FIG. 32A and FIG. 32B for preparing TetraGalNAc-siRNA Conjugate 19-1.

FIGS. 33A-33B. Scheme 35 as shown in FIG. 33A and FIG. 33B for preparing Compound 26.

FIGS. 34A-34C. Scheme 36 as shown in FIG. 34A to FIG. 34C for preparing Compounds 27 and 28.

FIGS. 35A-35B. Scheme 38 as shown in FIG. 35A and FIG. 35B for preparing Conjugates 35-37.

FIGS. 36A-36C. Scheme 39 as shown in FIG. 36A to FIG. 36C for preparing Conjugates 38-44.

FIG. 37. Scheme 40 as shown in FIG. 37 showing examples of different linkers from Table 2, for conjugating tetraGalNAc to siRNA.

FIGS. 38A-38E. Scheme 41 as shown in FIG. 38A to FIG. 38E for preparing Compounds and/or Conjugates 46-48.

FIGS. 39A-39C. Scheme 42 as shown in FIG. 39A to FIG. 39C for preparing Compounds and/or Conjugates 49-51.

FIG. 40. Scheme 43 as shown in FIG. 40 showing a general description for illustrative purposes of nomenclature used in Table 6.

DETAILED DESCRIPTION OF THE INVENTION

Disclosed herein are single chemical conjugates comprising a single stranded or double stranded oligonucleotide; one or more tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different;

wherein X is —O—, —S—, —CR¹R²— or —NR¹—, wherein R¹ and R² are each independently selected from the group consisting of hydrogen and C1-C6alkyl; n is 1, 2, 3, or 4; and the bond with “

” indicates the point of attachment; and one or more peptides, which may be the same or different. Other functionalities, such as targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents are optionally present. In one embodiment, R¹ and R² are each independently selected from the group consisting of hydrogen, methyl and ethyl. In another embodiment, R¹ and R² are each hydrogen.

In one embodiment, the oligonucleotide is a short interfering RNA (siRNA). In another embodiment, the siRNA is a single stranded siRNA. In another embodiment, the siRNA is a double stranded siRNA.

The use of the tetraGalNAc disclosed herein provides effective delivery of the oligonucleotide or siRNA by directing the modular composition to a particular cell. For example, the targeting ligand may specifically or non-specifically bind with a molecule on the surface of a target cell and facilitate internalization of the ligand-siRNA conjugate.

The peptides may function as endosomolytic, cell penetrating and/or fusogenic agents. In addition, the peptide may have cationic, zwitterionic, neutral, anionic character. Incorporation of both the tetraGalNAc and the peptide in the modular composition may further improve the delivery efficiency of the oligonucleotide or siRNA.

A linker may be present between each peptide and the oligonucleotide or between each tetraGalNAc and the oligonucleotide. The linkers are attached to the oligonucleotide at different 2′-positions of the ribose rings and/or the terminal 3′ and/or 5′-positions of the oligonucleotide.

In one embodiment, a modular composition comprises 1) a single stranded or double stranded oligonucleotide; 2) one or more tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different, wherein X is —O—, —S—, —CH₂— or —NH—; n is 1, 2, 3, or 4; and the bond with “

” indicates the point of attachment; optionally, 3) one or more linkers, which may be the same or different; 4) one or more peptides independently selected from Table 3, which may be the same or different; and optionally, 5) one or more targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.

In another embodiment, a modular composition comprises 1) a single stranded or double stranded oligonucleotide; 2) 1-8 tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different, wherein X is —O—, —S—, —CH₂— or —NH—; n is 1, 2, 3, or 4; 3) 1-24 linkers, which may be the same or different; 4) 1-8 peptides independently selected from Table 3, which may be the same or different; and optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.

In another embodiment, a modular composition comprises 1) a single stranded or double stranded siRNA; 2) 1-8 tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different, wherein X is —O—, —S—, —CH₂— or —NH—; n is 1, 2, 3, or 4; 3) 1-24 linkers, which may be the same or different; 4) 1-8 peptides independently selected from Table 3, which may be the same or different; and optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.

In one subset of the above embodiments, the tetraGalNAc ligands and/or the peptides are attached to the oligonucleotide or siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the oligonucleotide or siRNA.

In another subset of the above embodiments, the tetraGalNAc ligands and/or the peptides are attached to the oligonucleotide or siRNA optionally via linkers. In one embodiment, the linkers are present.

In another subset of the above embodiments, the tetraGalNAc ligands and/or the peptides are attached to the oligonucleotide or siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the oligonucleotide or siRNA; and the tetraGalNAc ligands and/or the peptides are attached to the oligonucleotide or siRNA via linkers.

In another subset of the above embodiments, the tetraGalNAc ligands are attached to the oligonucleotide or siRNA via linkers and the linkers are attached to the oligonucleotide or siRNA at different 2′-positions of the ribose rings.

In another subset of the above embodiments, the tetraGalNAc ligands are attached to the oligonucleotide or siRNA via linkers and the linkers are attached to the oligonucleotide or siRNA at different terminal 3′ and/or 5′-positions of the oligonucleotide.

In another subset of the above embodiments, X is —O—, —S—, or —CH₂—. In another embodiment, X is —O— or —CH₂—. In another embodiment, n is 1, 2 or 3. In another embodiment, X is —O— and n is 1 or 2. In another embodiment, X is —CH₂— and n is 1 or 2. In another embodiment, X is —O— and n is 1. In yet another embodiment, X is —CH₂— and n is 1.

In another subset of the above embodiments, the oligonucleotide or siRNA is single stranded. In another embodiment, the oligonucleotide or siRNA is double stranded.

In another subset of the above embodiments, the composition comprises 1-6 tetraGalNAc ligands. In another embodiment, the composition comprises 1-4 tetraGalNAc ligands. In another embodiment, the composition comprises 1-2 tetraGalNAc ligands. In yet another embodiment, the composition comprises 1 tetraGalNAc ligand.

In another subset of the above embodiments, the composition comprises 1-6 peptides. In another embodiment, the composition comprises 1-4 peptides. In another embodiment, the composition comprises 1-2 peptides. In yet another embodiment, the composition comprises 1 peptide.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the tetraGalNAc ligands are attached to the guide strand at different 2′-positions of the ribose rings.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded the tetraGalNAc ligands are attached to the guide strand at different terminal 3′ and/or 5′-positions.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the tetraGalNAc ligands are attached to the passenger strand at different 2′-positions of the ribose rings.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the tetraGalNAc ligands are attached to the passenger strand at different terminal 3′ and/or 5′-positions.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the tetraGalNAc ligands are attached to both the guide strand and the passenger strand at different 2′-positions of the ribose rings and/or different terminal 3′ and/or 5′-positions.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the peptides are attached to the guide strand at different 2′-positions of the ribose rings.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the peptides are attached to the guide strand at different terminal 3′ and/or 5′-positions.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the peptides are attached to the passenger strand at different 2′-positions of the ribose rings.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the peptides are attached to the passenger strand at different terminal 3′ and/or 5′-positions.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the peptides are attached to both the guide strand and the passenger strand at different 2′-positions of the ribose rings and/or different terminal 3′ and/or 5′-positions.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the tetraGalNAc ligands and the peptides are attached to the same or different strands via linkers. In one embodiment, each linker is independently selected Table 1.

In another embodiment, each linker is independently selected Table 2.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the tetraGalNAc ligands and the peptides are attached to the same strand.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the tetraGalNAc ligands and the peptides are attached to different strands.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the optional targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents are attached to the same or different strands.

In another subset of the above embodiments, the oligonucleotide or siRNA is double stranded and the optional targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents are attached to the same or different strands via linkers. In one embodiment, each linker is independently selected from Table 1. In another embodiment, each linker is independently selected from Table 2.

In one embodiment, a modular composition comprises 1) a single stranded or double stranded siRNA; 2) 1-8 tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different; wherein X is —O—, —S—, —CH₂— or —NH—; and n is 1, 2, 3, or 4; 3) 1-24 linkers, which may be the same or different; 4) 1-8 peptides independently selected from Table 3, which may be the same or different; and optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents; wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA optionally via linkers. In one embodiment, the linkers are present. In another embodiment, X is —O—, —S—, or —CH₂—, and n is 1, 2 or 3. In another embodiment, X is —O— or —CH₂—, and n is 1 or 2.

In another embodiment, a modular composition comprises 1) a double stranded siRNA; 2) 1-6 tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different; wherein X is —O—, —S—, or —CH₂—; and n is 1, 2 or 3; 3) 1-18 linkers, which may be the same or different; 4) 1-6 peptides independently selected from Table 3, which may be the same or different; and optionally, 5) 1-6 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents; wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA optionally via linkers. In one embodiment, the linkers are present. In another embodiment, X is —O—, —S—, or —CH₂— and n is 1 or 2. In another embodiment, the linkers are independently selected from Table 1. In another embodiment, the linkers are independently selected from Table 2. In another embodiment, the peptides of 4) are independently selected from Table 4.

In another embodiment, a modular composition comprises 1) a double stranded siRNA; 2) 1-4 tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different; wherein X is —O—, —S—, or —CH₂—; and n is 1 or 2; 3) 1-12 linkers, which may be the same or different; 4) 1-4 peptides independently selected from Table 3, which may be the same or different; and optionally, 5) 1-4 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents; wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA via linkers. In one embodiment, X is —O— or —CH₂— and n is 1 or 2. In another embodiment, the linkers are independently selected from Table 1. In another embodiment, the linkers are independently selected from Table 2. In another embodiment, the peptides are independently selected from Table 4.

In another embodiment, a modular composition comprises 1) a double stranded siRNA; 2) 1-4 tetraGalNAc ligands of Formula (IV), (V) or (VI), which may be the same or different:

3) 1-12 linkers independently selected from Table 1, which may be the same or different; 4) 1-4 peptides independently selected from Table 3, which may be the same or different; and optionally, 5) 1-4 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents; wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA via linkers.

In another embodiment, a modular composition comprises 1) a double stranded siRNA; 2) 1-4 tetraGalNAc ligands of Formula (IV), (V) or (VI); 3) 1-12 linkers independently selected from Table 2, which may be the same or different; 4) 1-4 peptides independently selected from Table 4, which may be the same or different; and optionally, 5) 1-4 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents; wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA via linkers.

In one subset of the above embodiments, the tetraGalNAc ligands and/or the peptides are attached to the siRNA via linkers; and wherein the tetraGalNAc ligands and/or the peptides are attached to the same strand.

In another subset of the above embodiments, the tetraGalNAc ligands and/or the peptides are attached to the siRNA via linkers; and wherein the tetraGalNAc ligands and the peptides are attached to different strands.

To illustrate the invention via cartoon, the invention features a modular composition, comprising an oligonucleotide ([O₁][O₂][O₃] . . . [O_(n)]), one or more tetraGalNAc(s) ligands (G), one or more linker(s) (L), one or more peptide(s) (P), and one or more optional lipid(s) (X), one or more targeting ligand(s) (X), and/or one or more solubilizing group(s) (X).

In an embodiment, the modular composition may have the formula:

G-L-[O₁][O₂][O₃] . . . [O_(n)]-L-P.

In another embodiment, the modular composition may have the formula:

P-L-[O1][O₂][O₃] . . . [O_(n)]-L-G.

Non-limiting examples of modular compositions comprising double stranded oligonucleotides with terminal conjugations are shown in FIG. 1.

Non-limiting examples of modular compositions comprising double stranded oligonucleotides with terminal conjugations are shown in FIG. 2.

Non-limiting examples of modular compositions comprising double stranded oligonucleotides with internal and/or terminal conjugations are shown in FIG. 3A and FIG. 3B.

These examples are used as illustration only. One skilled in the art will recognize that a variety of permutations for placing the desired components on the passenger and guide strand exist.

Any number of linkers, and therefore any number of peptides, can be attached to the oligonucleotide. The range of numbers of linkers is from 1-16. A more preferred range of numbers of linkers is from 1-12, or more specifically, 1-8, or even more specifically, 1-4.

The range of numbers of tetraGalNAc ligands is from 1-8. A more preferred range of numbers of tetraGalNAc ligands is from 1-6, or more specifically, 1-4, or even more specifically, 1-2.

The range of numbers of peptides is from 1-8. A more preferred range of numbers of peptides is from 1-6, or more specifically, 1-4, or even more specifically, 1-2.

The two strands contain n and n′ nucleotides respectively. The numbers n and n′ can be equal or different. The numbers are integers ranging from 8 to 50. Preferably, the numbers are integers ranging from 12-28. More preferably, the numbers are integers ranging from 19-21.

As an example, each nucleotide [O_(n)] or [O_(n).], that contains a linker (L-P and/or L-G) has generic structures as shown in FIG. 4.

For each nucleotide, 1) E=oxygen (O) or sulfur (S); 2) Base=A, U, G or C, which can be modified or unmodified; 3) D is the connection point between ribose ring and linker L, D=oxygen (O), sulfur (S, S(O) or S(O)₂), nitrogen (N—R, wherein R=H, alkyl, L-P or L-X), carbon (CH—R, wherein R=H, alkyl, L-P, or L-X), or phosphorus (P(O)R or P(O)(OR), wherein R=alkyl, L-P, or L-X). Preferably, D=oxygen (O).

The two nucleotides [O_(n-1)] and [O_(n)] or [O_(n-1)] and [O_(n)] are connected via phosphodiester or thio-phosphodiester bonds.

When the oligonucleotide is a double-stranded oligonucleotide, the “G-L”, “P-L” and the lipid, targeting ligand, and/or solubilizing group may be located on the same strand or on different strands.

In some embodiments, the “G-L” and “P-L” are on the same strand.

In some embodiments, the “G-L” and “P-L” are on the passenger strand.

In some embodiments, the “G-L” and “P-L” are on the guide strand.

In some embodiments, the “G-L” and “P-L” are located on different strands.

In some embodiments, the “G-L” is on the passenger strand while the “P-L” is on the guide strand.

In some embodiments, the “G-L” and “P-L” are on different strands but on the same terminal end of the double-stranded oligonucleotide.

In some embodiments, the “G-L” and “P-L” are on different strands and on the opposite terminal ends of the double-stranded oligonucleotide.

In some embodiments, the “G-L” can be located on multiple terminal ends of either the passenger or guide strand and “P-L” can be located on the remaining terminal ends of the passenger and guide strands.

In some embodiments, one “G-L” and two or more “P-L” are present in the oligonucleotide.

In some embodiments, two or more “G-L” and two or more “P-L” are present in the oligonucleotide.

In some embodiments, when the oligonucleotide is a double-stranded oligonucleotide and multiple “G-L” and/or “P-L” are present, such multiple “G-L” components and/or “P-L” may all be present in one strand or both strands of the double stranded oligonucleotide.

When multiple “G-L” components and/or “P-L” are present, they may all be the same or different.

In some embodiments, the “G-L” and/or “P-L” are on internal nucleotides only (i.e. excluding the 3′- and 5′-terminal ends of the oligonucleotide).

In another aspect, the invention includes a method of delivering an oligonucleotide or siRNA to a cell. The method includes (a) providing or obtaining a modular composition disclosed herein; (b) contacting a cell with the modular composition; and (c) allowing the cell to internalize the modular composition.

The method can be performed in vitro, ex vivo or in vivo, e.g., to treat a subject identified as being in need of an oligonucleotide or siRNA. A subject in need of said oligonucleotide is a subject, e.g., a human, in need of having the expression of a gene or genes, e.g., a gene related to a disorder, downregulated or silenced.

In one aspect, the invention provides a method for inhibiting the expression of one or more genes. The method comprising contacting one or more cells with an effective amount of an oligonucleotide of the invention, wherein the effective amount is an amount that suppresses the expression of the one or more genes. The method can be performed in vitro, ex vivo or in vivo.

The methods and compositions of the invention, e.g., the modular composition described herein, can be used with any oligonucleotides or siRNAs known in the art. In addition, the methods and compositions of the invention can be used for the treatment of any disease or disorder known in the art, and for the treatment of any subject, e.g., any animal, any mammal, such as any human. One of ordinary skill in the art will also recognize that the methods and compositions of the invention may be used for the treatment of any disease that would benefit from downregulating or silencing a gene or genes.

The methods and compositions of the invention, e.g., the modular composition described herein, may be used with any dosage and/or formulation described herein, or any dosage or formulation known in the art. In addition to the routes of administration described herein, a person skilled in the art will also appreciate that other routes of administration may be used to administer the modular composition of the invention.

Oligonucleotide

An “oligonucleotide” as used herein, is a double stranded or single stranded, unmodified or modified RNA or DNA. Examples of modified RNAs include those which have greater resistance to nuclease degradation than do unmodified RNAs. Further examples include those which have a 2′ sugar modification, a base modification, a modification in a single strand overhang, for example a 3′ single strand overhang, or, particularly if single stranded, a 5′ modification which includes one or more phosphate groups or one or more analogs of a phosphate group. Examples and a further description of oligonucleotides can be found in WO2009/126933, which is hereby incorporated by reference.

In an embodiment, an oligonucleotide is an antisense, miRNA, peptide nucleic acid (PNA), poly-morpholino (PMO) or siRNA. The preferred oligonucleotide is an siRNA. Another preferred oligonucleotide is the passenger strand of an siRNA. Another preferred oligonucleotide is the guide strand of an siRNA.

siRNA

siRNA directs the sequence-specific silencing of mRNA through a process known as RNA interference (RNAi). The process occurs in a wide variety of organisms, including mammals and other vertebrates. Methods for preparing and administering siRNA and their use for specifically inactivating gene function are known. siRNA includes modified and unmodified siRNA. Examples and a further description of siRNA can be found in WO2009/126933, which is hereby incorporated by reference.

A number of exemplary routes of delivery are known that can be used to administer siRNA to a subject. In addition, the siRNA can be formulated according to any exemplary method known in the art. Examples and a further discription of siRNA formulation and administration can be found in WO2009/126933, which is hereby incorporated by reference.

The phrases “short interfering nucleic acid”, “siNA”, “short interfering RNA”, “siRNA”, “short interfering nucleic acid molecule”, “oligonucleotide”, “short interfering oligonucleotide molecule”, or “chemically modified short interfering nucleic acid molecule” refer to any nucleic acid molecule capable of inhibiting or down regulating gene expression or viral replication by mediating RNA interference (“RNAi”) or gene silencing in a sequence-specific manner. These terms can refer to both individual nucleic acid molecules, a plurality of such nucleic acid molecules, or pools of such nucleic acid molecules. The siNA can be a double-stranded nucleic acid molecule comprising self-complementary sense and antisense strands, wherein the antisense strand comprises a nucleotide sequence that is complementary to a nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense strand comprises a nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. The siNA can be a polynucleotide with a duplex, asymmetric duplex, hairpin or asymmetric hairpin secondary structure, having self-complementary sense and antisense regions, wherein the antisense region comprises a nucleotide sequence that is complementary to a nucleotide sequence in a separate target nucleic acid molecule or a portion thereof and the sense region comprises a nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. The siNA can be a circular single-stranded polynucleotide having two or more loop structures and a stem comprising self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to a nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense region comprises a nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof, and wherein the circular polynucleotide can be processed either in vivo or in vitro to generate an active siNA molecule capable of mediating RNAi. The siNA can also comprise a single-stranded polynucleotide having a nucleotide sequence complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof (for example, where such siNA molecule does not require the presence within the siNA molecule of a nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof), wherein the single-stranded polynucleotide can further comprise a terminal phosphate group, such as a 5′-phosphate (see for example, Martinez et al., 2002, Cell, 110, 563-574 and Schwarz et al., 2002, Molecular Cell, 10, 537-568), or 5′,3′-diphosphate.

siRNA directs the sequence-specific silencing of mRNA through a process known as RNA interference (RNAi). The process occurs in a wide variety of organisms, including mammals and other vertebrates. Methods for preparing and administering siRNA and their use for specifically inactivating gene function are known. As used herein, siRNA includes chemically modified and unmodified nucleic acid molecules capable of inhibiting or down regulating gene expressions. Examples and a further discription of siRNA can be found in WO2009/126933, which is hereby incorporated by reference.

A number of exemplary routes of delivery are known that can be used to administer siRNA to a subject. In addition, the siRNA can be formulated according to any exemplary method known in the art. Examples and a further discription of siRNA formulation and administration can be found in WO2009/126933, which is hereby incorporated by reference.

Linkers

The covalent linkages between the tetraGalNAc and the oligonucleotide or siRNA of the modular composition and/or between the peptide and the oligonucleotide or siRNA may be mediated by a linker. This linker may be cleavable or non-cleavable, depending on the application. In certain embodiments, a cleavable linker may be used to release the oligonucleotide after transport from the endosome to the cytoplasm. The intended nature of the conjugation or coupling interaction, or the desired biological effect, will determine the choice of linker group. Linker groups may be combined or branched to provide more complex architectures. Suitable linkers include those as described in WO2009/126933, which is hereby incorporated by reference.

In one embodiment, the linkers of the instant invention are shown in Table 1:

TABLE 1

R = H, Boc, Cbz, Ac, PEG, lipid, targeting ligand, linker(s) and/or peptide(s). n = 0 to 750. “nucleotide” can be substituted with non-nucleotide moiety such as abasic or linkers as are generally known in the art. enzymatically cleavable linker = linker cleaved by enzyme; e.g., protease or glycosidase

In another embodiment, the preferred linkers are shown in Table 2.

TABLE 2

R = H, Boc, Cbz, Ac, PEG, lipid, targeting ligand, linker(s) and/or peptide(s). n = 0 to 750. “nucleotide” can be substituted with non-nucleotide moiety such as abasic or linkers as are generally known in the art. enzymatically cleavable linker = linker cleaved by enzyme; e.g., protease or glycosidase

Commercial linkers are available from various suppliers such as Pierce or Quanta Biodesign including combinations of said linkers. In addition, commercial linkers attached via phosphate bonds can be used independently as linkers or in combination with said linkers. The linkers may also be combined to produce more complex branched architectures accomodating from 1 to 8 peptides as illustrated in one such example below:

Peptides

For macromolecular drugs and hydrophilic drug molecules, which cannot easily cross bilayer membranes, entrapment in endosomal/lysosomal compartments of the cell is thought to be the biggest hurdle for effective delivery to their site of action. Without wishing to be bound by theory, it is believed that the use of peptides will facilitate oligonucleotide escape from these endosomal/lysosomal compartments or oligonucleotide translocation across a cellular membrane and release into the cytosolic compartment. In certain embodiments, the peptides of the present invention may be polycationic or amphiphilic or polyanionic or zwitterionic or lipophilic or neutral peptides or peptidomimetics which can show pH-dependent membrane activity and/or fusogenicity. A peptidomimetic may be a small protein-like chain designed to mimic a peptide.

In some embodiments, the peptide is a cell-permeation agent, preferably a helical cell-permeation agent. These peptides are commonly referred to as Cell Penetrating Peptides. See, for example, “Handbook of Cell Penetrating Peptides” Ed. Langel, U.; 2007, CRC Press, Boca Raton, Fla. Preferably, the component is amphipathic. The helical agent is preferably an alpha-helical agent, which preferably has a lipophilic and a lipophobic phase. A cell-permeation agent can be, for example, a cell permeation peptide, cationic peptide, amphipathic peptide or hydrophobic peptide, e.g. consisting primarily of Tyr, Trp and Phe, dendrimer peptide, constrained peptide or crosslinked peptide. Examples of cell penetrating peptides include Tat, Penetratin, and MPG. For the present invention, it is believed that the cell penetrating peptides can be a “delivery” peptide, which can carry large polar molecules including peptides, oligonucleotides, and proteins across cell membranes. Cell permeation peptides can be linear or cyclic, and include D-amino acids, “retro-inverso” sequences, nonpeptide or pseudo-peptide linkages, peptidyl mimics. In addition the peptide and peptide mimics can be modified, e.g. glycosylated, pegylated, or methylated. Examples and a further discription of peptides can be found in WO2009/126933, which is hereby incorporated by reference. Synthesis of peptides is well known in the art.

The peptides may be conjugated at either end or both ends by addition of a cysteine or other thiol containing moiety to the C- or N-terminus. When not functionalized on the N-terminus, peptides may be capped by an acetyl group, or may be capped with a lipid, a PEG, or a targeting moiety. When the C-terminus of the peptides is unconjugated or unfunctionalized, it may be capped as an amide, or may be capped with a lipid, a PEG, or a targeting moiety.

Suitable peptides that can be used in the conjugates disclosed herein are listed in Table 3 below:

TABLE 3 Peptide Sequence Listing and ID Sequence SEQ ID CGLFEAIEEFIENLWELLIDGWYGYGRKKRRQRR SEQ ID NO: 1 CGLFEAIEGFIENGWEGMIDGWYGYGHKKHHQHH SEQ ID NO: 2 C-bAla-LFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 3 CGLFEAIEGFIENGLKGLIDWWYGYGRKKRRQRR SEQ ID NO: 4 CGLFEAIEGFIEWGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 5 CRRQRRKKRGYGYWGDIMGEWGNEIFGEIAEFLG SEQ ID NO: 6 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQR SEQ ID NO: 7 CYGRKKRRQRRGLFEAIEGFIENGWEGMIDGWYG SEQ ID NO: 8 CIFGAIAGFIKNILKGLIDG SEQ ID NO: 9 CIFGAIAGFIRNIW SEQ ID NO: 10 CGLFHALLHLLHSLWHGLLHAWYGYGHKKHHQHR SEQ ID NO: 11 CGLFEAIEGLIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 12 CGLFELIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 13 CGLFEAIEGFIENGWEGLIDGWYGYGOOOOOQRR (O = ornithine) SEQ ID NO: 14 CGLFGAIEGFIENGWEGLIDGWYGYGRKKRRQRR SEQ ID NO: 15 CGLFEAIEGFLENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 16 CGLFEAIEGFIENGLEGMIDGWYGYGRKKRRQRR SEQ ID NO: 17 CGLFGAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 18 CGLFEAIEGFIENGWEG-Nle-IDGEYGYGRKKRRQRR SEQ ID NO: 19 CGIFGAIAGFIKNIWKGLIDW SEQ ID NO: 20 CYGRKKRRQRRGLFEAIEGFIENGWKGLIDAWYG SEQ ID NO: 21 CGLLEALEGLLESLWEGLLEAEYGYGRKKRRQRR SEQ ID NO: 22 CGLFEAIEGFIENGWEGMIDNEYGYGRKKRRQRR SEQ ID NO: 23 CIFGAIAGFIKNIWEGLIEAWYGLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 24 CIFGAIAGFIKNIWEGLIDAF SEQ ID NO: 25 CIFGAIAGFIKNIWEGLI SEQ ID NO: 26 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRK (stearyl) SEQ ID NO: 27 CGLFEAIAGFIEGGWPGLINGWYGYGRKKRRQRRLHLLHHLLHHLHHLLHHLLHLL SEQ ID NO: 28 HHLLHHL CGLFEAIEGFIENGWEGMIDGWYGGGGLHLLHHLLHHLHHLLHHLLHLLHHLLHHL SEQ ID NO: 29 CGLFEAIEGFIENGWEGMIDGWYGLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 30 CGLFEALLELLESLWELLLEAYGRKKRRQRR SEQ ID NO: 31 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 32 CGLFEAIEGFIENGWEGMADGWYGYGRKKRRQRR SEQ ID NO: 33 CGIFGAIAGFIKNIWEGLIDWWYGYGRKKRRQRR SEQ ID NO: 34 CGFLPAIAGILSQLFEGLIDGWYGYGRKKRRQRR SEQ ID NO: 35 CFFGAIWGFIKSIL SEQ ID NO: 36 CIFGAIAGFIKNIWKGLIDWWYG SEQ ID NO: 37 CGLFEAIEGFIWNGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 38 CGLFEAIAEFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 39 CYGRKKRRQRRGLFEAIEGFIENGWKGLIDWWYG SEQ ID NO: 40 CGLFEAIEGFIEEGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 41 CGLFEAIEGFIENAWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 42 CGLFEAIEGFIENGWEGMIDLWYGYGRKKRRQRR SEQ ID NO: 43 CRLLRLLLRLWRRLLRLLR SEQ ID NO: 44 CGGFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 45 CGLFEKIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 46 CGLFEAIEGFIENGWENMIDGWYGYGRKKRRQRR SEQ ID NO: 47 CIFGAIAGFIKNILKGL SEQ ID NO: 48 CIFGAIAGFIKNILKGLIDGWYG SEQ ID NO: 49 CGLFEAIEGFIENGWEGMIDGWYG-(PEG)3-YGRKKRRQRR SEQ ID NO: 50 CGLFEALLELLESLWELLLEAYGRKKRRQRRLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 51 CYGRKKRRQRRWEAALAEALAEALAEHLAEALAEALEALAA SEQ ID NO: 52 CIFGAIAGFIKNIWEGLIDGWYGKLALKLALKALKAALKLA SEQ ID NO: 53 CFFGAIWEFIRSILEGLIDGWYGYGRKKRRQRR SEQ ID NO: 54 CGLFHALLHLLHSLWHLLLHAWYGYGRKKRRQRR SEQ ID NO: 55 CGLFHALLHLLHSLWHLLLHAWYGYGHKKHHQHR SEQ ID NO: 56 CGLFGALLELLESLWKGLLEWYGRKKRRQRR SEQ ID NO: 57 CRRQRRKKRGYGYWGDILGEWGNEIFGEIAEFLG SEQ ID NO: 58 CGLFEALEGFLENGWEGLLDGWYGYGROORRQRR (O = ornithine) SEQ ID NO: 59 CGLFGEIEELIENGLKNLIDWWYGYGRKKRRQRR SEQ ID NO: 60 CRRQRRKKRGYGYWWDILGKWGNEIFGEIAEFLG all (D) aminos SEQ ID NO: 61 CGIFGAIAGFIKNIL SEQ ID NO: 62 CGIFGAIAGLLKNIFK SEQ ID NO: 63 CIFGAIAGFIKNIWKGLIDW SEQ ID NO: 64 CIFGAIAGFIKNIWK SEQ ID NO: 65 CGLFEEIEGFIENGWEGLIDWWYGYGHKKHHQHR SEQ ID NO: 66 CGLFGEIEELIENGLKNLIDWWYGYGHKKHHQHR SEQ ID NO: 67 CGLFEEIEEFIENGWEGLIDWWYGYGHKKHHQHR SEQ ID NO: 68 stearyl-WEAALAEALAEALAEHLAEALAEALEALAAYGRKKRRQRRC SEQ ID NO: 69 CGLFEAIEGFIENGWKGLIDGWYGGLFEAIEGFIENGWKGLIDWWYG SEQ ID NO: 70 CGFFHAFFHFFHSFWHGFFEA SEQ ID NO: 71 CGNFGEIEELIEEGLENLIDWWNG SEQ ID NO: 72 CFFGAIWEFIRNILEGF SEQ ID NO: 73 CFFGAIWEFIHSIL SEQ ID NO: 74 CGLFHALLHLLHSLWHGLLEA SEQ ID NO: 75 CIFGAIAGFIKNIWEGL SEQ ID NO: 76 CIFGAIAGLLKNIFEGLIDGWYGYGRKKRRQRR SEQ ID NO: 77 CGFIGAIANLLSKIFEGLIDGWWYGYGRKKRRQRR SEQ ID NO: 78 CGLFEAIEELIENLWKGLIDAWWYGYGRKKRRQRR SEQ ID NO: 79 CGIFGAIAGLLKNIFKGLIDA SEQ ID NO: 80 CGIFGAIAGLLKNIFKGLIDW SEQ ID NO: 81 CGIFEAIAGLLKNIFK SEQ ID NO: 82 CGIFEEIAGLLKNIFK SEQ ID NO: 83 CGLFEAIAGFIEGGWPGLINGWYGYGRKKRRQRRLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 84 CGLFEAIEGFIENGWKGMIDWWYGYGRKKRRQRRK (stearyl) SEQ ID NO: 85 CGLFGEIEEFIENGWKGLIDWWYG SEQ ID NO: 86 CIFGAIAGFIKNIWLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 87 CGIFGAIEGFIENGWKGLIDAWYGYRKKRRQRR SEQ ID NO: 88 CELFGAIEGFIENGWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 89 CIFGIDDLIIGLLFVAIVEAGIGGYLLGSYGRKKRRQRR SEQ ID NO: 90 GLFGALAEALAEALAEHLAEALAEALEALAAGGSC SEQ ID NO: 91 CGFIGAIANLLSKIFEGLIDGWWYGYGRKKRRQRR all (D) SEQ ID NO: 92 CFFGAIWEFIRSILKGLI SEQ ID NO: 93 CFFGAIWEFIRSILK SEQ ID NO: 94 CFFGAIWEFIRSILE SEQ ID NO: 95 CIFGAIAGFIKNIWE SEQ ID NO: 96 CIFGAIAGFIKNIWKGLIDA SEQ ID NO: 97 CFFEAIEEFIKNILK SEQ ID NO: 98 CIFGAIAGLLRNIF SEQ ID NO: 99 CGIFGAIAGLLKNIW SEQ ID NO: 100 CLFGAIWEFIKSIL SEQ ID NO: 101 CFWGAIWEFIKSIL SEQ ID NO: 102 CFGGAIWEFIKSIL SEQ ID NO: 103 CFAGAIWEFIKSIL SEQ ID NO: 104 CGLFEAIEGFIENGWEGM(SO2)IDGWYGYGRKKRRQRR SEQ ID NO: 105 CGLFEAIEGFIENGWEGMIDWWYGYGRKKRRQRR SEQ ID NO: 106 CFFGAIWEFIKSIG SEQ ID NO: 107 CFFGAIWEFIKSIA SEQ ID NO: 108 CFFGAIWEFIKSIN SEQ ID NO: 109 CFFGAIWEFIKSIW SEQ ID NO: 110 CFFGAIWEFIKSILEGLIDWWYGYGHKKHHQHR SEQ ID NO: 111 Ac-CLHLLHHLLHHLHHLLHHLLHLLHHLLHHL-NH2 SEQ ID NO: 112 Ac-LHLLHHLLHHLHHLLHHLLHLLHHLLHHLGGGRKKRRQRRRPPQC-NH2 SEQ ID NO: 113 CRKKRRQRRRPPQGGGLHLLHHLLHHLHHLLHHLLHLLHHLLHHL SEQ ID NO: 114 CLHLLHHLLHHLHHLLHHLLHLLHHLLHHLGGGRKKRRQRRRPPQ SEQ ID NO: 115 CGLFHAIAHFIHGGWHGLIHGWWYGYGRKKRRQRR SEQ ID NO: 116 CGLFKAIAKFIKGGWKGLIKGWWYGYGRKKRRQRR SEQ ID NO: 117 CGLFEAIAGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 118 CWEAALAEALAEALAEHLAEALAEALEALAAYGRKKRRQRR SEQ ID NO: 119 CGLFEAIEGFIENGWEGMIDGWWYGRKKRRQRRRPPQ SEQ ID NO: 120 GLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRC SEQ ID NO: 121 Ac-LHLLHHLLHHLHHLLHHLLHLLHHLLHHLRKKRRQRRRPPQ-NH2 SEQ ID NO: 122 Ac-LHLLHHLLHHLHHLLHHLLHLLHHLLHHLGPGRKKRRQRRRPPQ-NH2 SEQ ID NO: 123 Ac-LIRLWSHLIHIWFQNRRLKWKKK-NH2 SEQ ID NO: 124 Ac-RKKRRQRRRPPQQQQQQ-NH2 SEQ ID NO: 125 Ac-GLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR-NH2 SEQ ID NO: 126 Ac-LHLLHHLLHHLHHLLHHLLHLLHHLLHHLGGGRRRRRRRRR-NH2 SEQ ID NO: 127 Ac-LHLLHHLLHHLHHLLHHLLHLLHHLLHHL-(Peg)12-RKKRRQRRRPPQ-NH2 SEQ ID NO: 128 Ac-GLFGAIAGFIENGWEGMIDGWWYGLIRLWSHLIWFQNRRLKWLLL-NH2 SEQ ID NO: 129 Ac-HHHHHRKKRRQRRRPPQGGGLHLLHHLLHHLHHLLHHLLHLLHHLLHHL-NH2 SEQ ID NO: 130 Ac-LHLLHHLLHHLHHLLHHLLHLLHHLLHHL-(Peg)2-RKKRRQRRRPPQ-NH2 SEQ ID NO: 131 Ac-LHLLHHLLHHLHHLLHHLLLLHHLLHHLGGGRQIKIWFQNRRMKWKKGG-NH2 SEQ ID NO: 132 Ac-KLLKLLLKLWLKLLKLLLKLLGGGRKKRRQRRRPPQ-NH2 SEQ ID NO: 133 Ac-LHHLLHHLLHLLHHLLHHLHHLLHHLLHLC-NH2 all (D) SEQ ID NO: 134 Ac-LHLLHHLLHHLHHLLHHLLHLLHHLLHHL-PEG6-RKKRRQRRRPPQC-NH2 SEQ ID NO: 135 Ac-GLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRC-NH2 SEQ ID NO: 136 CGLFEAIEGFIENGWEGMIDGWWYGYGRKKRRQRR all (D) SEQ ID NO: 137 CGLFEAIEGFIENGWEGMIDGWYGYGRRRRRRRRR-NH2 SEQ ID NO: 138 YGRKKRRQRRGLFEAIEGFIENGWEGMIDGWWYGC-NH2 SEQ ID NO: 139 CGVFVLGFLGFLATAGSYGRKKRRQRR-NH2 SEQ ID NO: 140 CGLFKAIAKFIKGGWKGLIKGWYG-NH2 SEQ ID NO: 141 CGLFEAIEGFIENGWEGMIDGWYGYGRKKR SEQ ID NO: 142 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRYGRKKRRQRR SEQ ID NO: 143 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRYGRKKRRQRR SEQ ID NO: 144 CGLFEAIKGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 145 CGLFEAIHGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 146 CGLFEAIRGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 147 CGLFEAIDGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 148 CRLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 149 CGGGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 150 CGLFEAIEGFIENGWEGMIDGWYGGGGYGRKKRRQRR SEQ ID NO: 151 CGLFEAIEGFIENGWEGMIDGWYG-(PEG)11-YGRKKRRQRR SEQ ID NO: 152 CFLGFLLGVGSAIASGIAVSKVLHL SEQ ID NO: 153 CGVFVLGFLGFLATAGSAMGARSLTLSAYGRKKRRQRR SEQ ID NO: 154 Ac-GLWRALWRLLRSLWRLLWRA-mercaptoethylamide SEQ ID NO: 155 C-Nle-LFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 156 CELFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 157 CGFFGAIAGFLEGGWEGMIAGWHGYGRKKRRQRR SEQ ID NO: 158 CFLGFLLGVGSAIASGIAVSKVLHLYGRKKRRQRR SEQ ID NO: 159 GLFEAIEGFIENGWEGLAEALAEALEALAAGGSC SEQ ID NO: 160 CGLFEAIEGFIENGWEGMIDGWYGLHLLHHLLHHLHHLLHHLLHLLHHLLHHL SEQ ID NO: 161 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRLHLLHHLLHHLHHLLHHLLHLL SEQ ID NO: 162 HHLLHHL CGLFGAIAGFIEGGWTGMIDGWYGYGRKKRRQRR SEQ ID NO: 163 CGLFGAIAGFIEGGWQGMVDGWYGYGRKKRRQRR SEQ ID NO: 164 CGLFGAIAGFIENGWQGLIDGWYGYGRKKRRQRR SEQ ID NO: 165 CGLFGAIAGFIENGWEGLVDGWYGYGRKKRRQRR SEQ ID NO: 166 CGLFGAIAGFIEGGWSGMIDGWYGYGRKKRRQRR SEQ ID NO: 167 CGLFGAIAGFIEGGWPGLVAGWYGYGRKKRRQRR SEQ ID NO: 168 CGLFGAIAGFIENGWEGMVDGWYGYGRKKRRQRR SEQ ID NO: 169 CGLFGAIAGFIEGGWPGLINGWYGYGRKKRRQRR SEQ ID NO: 170 CGLFGAIAGFIENGWEGLIDGWYGYGRKKRRQRR SEQ ID NO: 171 CGLFGAIAGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 172 CGLFGAIAGFIENGWEGMIDGWYGSSKKKK SEQ ID NO: 173 CGLFGAIAGFIENGWEGLIDGWYGYGRKKRRQRR SEQ ID NO: 174 CGLFEAIEGFIENGWEGLIDGWYGYGRKKRRQRR SEQ ID NO: 175 CGLFGAIAGFIENGWEGLIEGWYGGGRKKRRQRR SEQ ID NO: 176 CGLFEAIEGFIENGWEGMIDGWYGGGRKKRRQRR SEQ ID NO: 177 CGLFEAIAGFIENGWEGLIDGWYGYGRKKRRQRR SEQ ID NO: 178 CGLFEAIAEFIENGWEGLIEGWYGGRKKRRQRR SEQ ID NO: 179 CGLFEAIEGFIENGWEGMIDGWYGRKKRRQRRR SEQ ID NO: 180 CKLLKLLLKLWLKLLKLLLKLL SEQ ID NO: 181 CKLLKLLLKLWLKLLKLLLKLLYGRKKRRQRR SEQ ID NO: 182 GLFEAIEGFIENGWEGMIDGWYGC SEQ ID NO: 183 CVLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 184 CSLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 185 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQ SEQ ID NO: 186 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRR SEQ ID NO: 187 CGLFEAIEGFIENGWEGMIDGWYGYGKKKKKQKK SEQ ID NO: 188 CGLFEAIEGFIENGWEGMIDGWYGGLFEAIEGFIENGWEGMIDGWYG SEQ ID NO: 189 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRGLFEAIEGFIENGWEGMIDG SEQ ID NO: 190 WYGYGRKKRRQRR RRQRRKKRGYGYWGDIMGEWGNEIFGEIAEFLGC SEQ ID NO: 191 CRRQRRKKRGYGYWGDIMGEWGNEIFGEIAEFLG SEQ ID NO: 192 GLFEAIEGFIENGWEGMIDGWYGYGRK-K(D)-RRQRR SEQ ID NO: 193 GLFEAIEGFIENGWEGMIDGWYGYGRKK-R(D)-RQRR SEQ ID NO: 194 GL-F(D)-EAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 195 GLF-E(D)-AIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 196 CGLFEAIEGFIENGWEGMIDGWWYG SEQ ID NO: 197 CYGRKKRRQRR SEQ ID NO: 198 YGRKKRRQRRC SEQ ID NO: 199 RRQRRKKRGYGYWGDIMGEWGNEIFGEIAEFLGC all(D) SEQ ID NO: 200 CRRQRRKKRGYGYWGDIMGEWGNEIFGEIAEFLG all(D) SEQ ID NO: 201 CGLFEAIEGFIENGWEGMIDGAYGYGRKKRRQRR SEQ ID NO: 202 CGLFEALLELLESLWELLLEAWYGYGRKKRRQRR SEQ ID NO: 203 CGLFEAIEGFNENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 204 CGLFEAIEGFIENEWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 205 K(stearoyl)GLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRC SEQ ID NO: 206 CGLFEAIK(stearoyl)GFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 207 CGLFEAIKGFIENGWEGMIDGWYGYGRK(stearoyl)KRRQRR SEQ ID NO: 208 CGLFEAIEGFIENPWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 209 (stearyl)GLFEAIEGFIENPWEGMIDGWWYGYGRKKRRQRRC SEQ ID NO: 210 CGLFGAIAGFIEGGWPGLINGWWYGYGRKKRRQRRLHLLHHLLHHLHHLLHHLLHLL SEQ ID NO: 211 HHLLHHL CGLFGAIAGFIEGGWPGLINGWYGYGRKKRRQRRLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 212 CGLFEAIAGFIEGGWPGLINGWYGYGRKKRRQRR SEQ ID NO: 213 CGLEEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 214 CGLFNAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 215 CGLFAAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 216 CGLFEAIENFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 217 CGLFEAIEKFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 218 CGLFEAIEGFAENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 219 CGLFEAIEGFIENWWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 220 CGLFEAIEGFIENNWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 221 CGLFEAIEGFIENGEEGMIDGWYGYGRKKRRQRR SEQ ID NO: 222 CGLFEAIEGFIENGWAGMIDGWYGYGRKKRRQRR SEQ ID NO: 223 CGLFEAIEGFIENGWNGMIDGWYGYGRKKRRQRR SEQ ID NO: 224 CGLFEAIEGFIENGWGGMIDGWYGYGRKKRRQRR SEQ ID NO: 225 CGLFEAIEGFIENGWEGMIDAWYGYGRKKRRQRR SEQ ID NO: 226 CGLFEAIEGFIENGWLGMIDGWYGYGRKKRRQRR SEQ ID NO: 227 CGLFEAIEGFIENGWKGMIDGWYGYGRKKRRQRR SEQ ID NO: 228 CGLFEAIEGFIENGWEGMIDKWYGYGRKKRRQRR SEQ ID NO: 229 CGLFEAIEGFIENGWEGMIDEWYGYGRKKRRQRR SEQ ID NO: 230 CGLFEAIEGFIENGWEGMIDGLYGYGRKKRRQRR SEQ ID NO: 231 CGLFEAIEGFIENGWEGMIDGNYGYGRKKRRQRR SEQ ID NO: 232 CGLFEAIEGFIENGWEGMIDGKYGYGRKKRRQRR SEQ ID NO: 233 CGLFEAIEGFIENGWEGMIDGEYGYGRKKRRQRR SEQ ID NO: 234 CGLFEALEELLEGGWEGLIEAWYGYGRKKRRQRR SEQ ID NO: 235 CELFGAIWEFIEGGWEGLIEAWYGYGRKKRRQRR SEQ ID NO: 236 CGLFEALEEFIEGGWEGLLEAWYGYGRKKRRQRR SEQ ID NO: 237 CGLFEALEEFIENGWEGLLEAWYGYGRKKRRQRR SEQ ID NO: 238 CGLFEAIEGFIESGWEGLIDGWYGYGRKKRRQRR SEQ ID NO: 239 CGLFEAIEEFIEGGWEGLIEAWYGYGRKKRRQRR SEQ ID NO: 240 CGLFEAIEGFIENGWEGLIDAWYGYGRKKRRQRR SEQ ID NO: 241 CGLFEAIEGFILNGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 242 CGLFEAIEGFIKNGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 243 CGLFEAIEGFIGNGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 244 CGLFEAIEGFIELGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 245 CGLFEAIEGFIEKGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 246 CGLFEAIAEFIEGGWEGLIEGWYGYGRKKRRQRR SEQ ID NO: 247 CRGWEVLKYWWNLLQY SEQ ID NO: 248 CRGWEVLKYWWNLLQYYGRKKRRQRR SEQ ID NO: 249 CGLFGAIAGFIENGWEGMIDGWYGFRYGRKKRRQRR SEQ ID NO: 250 Ac-CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR-CO2H SEQ ID NO: 251 CGLLEALEGLLENGWEGLLEAWYGYGRKKRRQRR SEQ ID NO: 252 CLRHLLRHLLRHLRHLLRHLRHLLRHLLRH SEQ ID NO: 253 CGIFEAIEGFIENGWEGIIDGWYGYGROORRQRR (O = ornithine) SEQ ID NO: 254 CGIGAVLKVLTTGLPALISWIKRKRQQ SEQ ID NO: 255 CGIGAVLKVLTTGLPALISWIHHHHQQ SEQ ID NO: 256 CGAFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 257 Ac-LHLLHHLLHHLHHLLHHLLHLLHHLLHHLRRRRR SEQ ID NO: 258 CGLFGAIWGFIENWWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 259 CGLFGAIEGFIENGWKGLIDAWYGYGRKKRRQRR SEQ ID NO: 260 CGLFEAIAGFIENGWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 261 GLFEAIEGFIENGWKGLIDWWYGYGRKKRRQRRC SEQ ID NO: 262 YGRKKRRQRRGLFEAIEGFIENGWKGLIDAWYGC SEQ ID NO: 263 YGRKKRRQRRGLFEAIEGFIENGWKGLIDWWYGC SEQ ID NO: 264 CGLFHAIHGFIENGWHGLIDWWYGYGRKKRRQRR SEQ ID NO: 265 CGLFEAIEGFIENGWKGLIDWWYGYGRKKRRQRRK (stearyl) SEQ ID NO: 266 CGLFKALLKLLKSLWKLLLKAWWYGYGHKKHHQHR SEQ ID NO: 267 CGLFKALLKLLKSLWKGLLKAWYGYGHKKHHQHR SEQ ID NO: 268 CGLAKALLKLLKSLWKGLIEAWWYGYGRKKRRQRR SEQ ID NO: 269 CGIFGAIAGFIKNIW SEQ ID NO: 270 CIFGAIAGFIKNIWEGLIDGWYGYGRKKRRQRR SEQ ID NO: 271 CGIFGAIAGFIKNIWEGLIDGYGRKKRRQRR SEQ ID NO: 272 CGIFGAIAGFIKNIWKGLIDAWWYGYGRKKRRQRR SEQ ID NO: 273 CIFGAIAGFIKNIWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 274 CLFGAIAGFIKNIW SEQ ID NO: 275 CGL(R5)EAIEGF(58)ENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 276 CGLFEA(55)EGF(55)ENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 277 CGLFEAIEGFIENGWEGAIDGWWYGYGRKKRRQRR SEQ ID NO: 278 CGLFEAIEGFIENGWEGEIDGWWYGYGRKKRRQRR SEQ ID NO: 279 CGIFGAIAGFIKNGWEGMVDWYGYGRKKRRQRR SEQ ID NO: 280 CGLFEAIAGFIENGWEGMIDGWYGFYGRKKRRQRR SEQ ID NO: 281 CGIFGAIAGFIKNGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 282 CIFGAIAGFIKNIW SEQ ID NO: 283 CIFGAIAGFIKNIWWYGRKKRRQRR SEQ ID NO: 284 CGIFGAIAGFIKNIWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 285 CGLFEAIEGFIENGWEGLIEAYGRKKRRQRR SEQ ID NO: 286 CGLFEALLGFIENGWEGLIDGYGRKKRRQRR SEQ ID NO: 287 CGLFGAIEGFIENGWEGLIDGWWYGYGRKKRRQRRR SEQ ID NO: 288 CELFGAIEGFIENGWEGMIDGWWYGYGRKKRRQRRR SEQ ID NO: 289 CGLFEAIEGFIENGWEGMIDGWYGYGHKKHHQHR SEQ ID NO: 290 CGLFGAIEGFIEGGWPGLINGWWYGYGRKKRRQRRR SEQ ID NO: 291 CGLFKALLKLLKSLWKLLLKAYGRKKRRQRR SEQ ID NO: 292 CGLFKALLKLLKSLWKLLLKAWYGYGRKKRRQRR SEQ ID NO: 293 CGLFRALLRLLRSLWRLLLRAYGRKKRRQRR SEQ ID NO: 294 CGLFEAILGFIENGWEGLIDGWYGYGRKKRRQRR SEQ ID NO: 295 CGLFEAIWEFIENGWEGLIDGWWYGYGRKKRRQRR SEQ ID NO: 296 CGLFEAIEGFIENGWEGMIDGWWYGGGGLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 297 CGPVEDAITAAIGRVADTVGTYGRKKRRQRR SEQ ID NO: 298 CMDGTLFPGDDDLAIPATEFFSTKA SEQ ID NO: 299 CGLFEALEEFIEGGWEGLLEAWYGYGRKKRRQRR SEQ ID NO: 300 CGLFEALEEFIENGWEGLLEAWYGYGRKKRRQRR SEQ ID NO: 301 CELFGAIWEFIEGGWEGLIEAYGRKKRRQRR SEQ ID NO: 302 CGLFEAIEGFIEEGWEGMIDGWWYGYGRKKRRQRR SEQ ID NO: 303 CGLFEAIAEFIENGWEGMIDGWWYGYGRKKRRQRR SEQ ID NO: 304 CGLFEAIAEFIEGLWEGLIEGWYGYGRKKRRQRR SEQ ID NO: 305 CGLLEALEGLLESLWEGLLEAWYGYGRKKRRQRR SEQ ID NO: 306 CGLFEAIEGFIENGWEGMIDIWYGYGRKKRRQRR SEQ ID NO: 307 CGLFEAIEGFIENGWRGMIDGWWYGYGRKKRRQRR SEQ ID NO: 308 CGLFEAIEGFIENGWDGMIDGWWYGYGRKKRRQRR SEQ ID NO: 309 CGLFEAIEGFIENHWEGMIDGWWYGYGRKKRRQRR SEQ ID NO: 310 CGLFEAIEGFIENWWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 311 GLFEAIEGFIENGWKGLIDAWWYGYGRKKRRQRRC SEQ ID NO: 312 CGLFEAIEGFIENGWKGMIDAWYGYGRKKRRQRR SEQ ID NO: 313 CGLFEAIEGFIENGWKGMIDWWYGYGRKKRRQRR SEQ ID NO: 314 CGLAEAIEGFIENGLKGLIDWWYGYGRKKRRQRR SEQ ID NO: 315 RRQRRKKRGYGYWGDILGEWGNEIFGEIAEFLGC all(D) SEQ ID NO: 316 CRRQRRKKRGYGYWWGDILGEWGNEIFGEIAEFLG all(D) SEQ ID NO: 317 CGLFEAIEGFIENGWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 318 CGFFEAIEGFIENGLKGLIDAWWYGYGRKKRRQRR SEQ ID NO: 319 CGLFEAIEGFIENGLKGLIDAWYGYGRKKRRQRR SEQ ID NO: 320 CELFGAIEGFIENGWKGLIDAWWYGYGRKKRRQRR SEQ ID NO: 321 CGLFKAIKGFIKNGWKGLIKAWWYGYGRKKRRQRR SEQ ID NO: 322 CGLAEALLELLESLWKGLIEAYGRKKRRQRR SEQ ID NO: 323 CGIFGAIEGFIENGWKGLIDAWYGYGRKKRRQRR SEQ ID NO: 324 CGIAGAIAGFIKNIWEGLIDWWYGYGRKKRRQRR SEQ ID NO: 325 CGIAGAIAGFIKNIWKGLIDAWYGYGRKKRRQRR SEQ ID NO: 326 CGIFGAIAGFIKNIWEGLIDGWWYGKKKKKKKKK SEQ ID NO: 327 CG(R5)FEAIEG(58)IENGWEGMIDGWWYGYGRKKRRQRR SEQ ID NO: 328 CGLFEAIEGF(R5)ENGWEG(S8)IDGWWYGYGRKKRRQRR SEQ ID NO: 329 GLFEAIEGFIENGWEGMIDGWYGCYGRKKRRQRR SEQ ID NO: 330 GLFEAIEGFIENGWEGMIDGWWYGGCGYGRKKRRQRR SEQ ID NO: 331 GLLEALEGLLENGWEGLLDGWYGYGRKKRRQRR SEQ ID NO: 332 CFFGAIWEFIRNIL SEQ ID NO: 333 CIFGAIAGFIRSIL SEQ ID NO: 334 CGLFEEIEEFIENGWEGLIDWWYGYGRKKRRQRR SEQ ID NO: 335 CGFFGAIWEFIKSIL SEQ ID NO: 336 GFFGAIWEFIKSILC SEQ ID NO: 337 CGLFEALEGFIENGWEGLLDGWWYGYGROORRQRR (O = ornithine) SEQ ID NO: 338 CGLFEALLELLENGWELLLEAWWYGYGRKKRRQRR SEQ ID NO: 339 CGLFEALLELLENGWELLLDGWYGYGRKKRRQRR SEQ ID NO: 340 CALFEAIEAFIENGWEAMIDAWYGYGRKKRRQRR SEQ ID NO: 341 CGLFGAIWGFIENGWEGLIDGWYGYGRKKRRQRR SEQ ID NO: 342 CGLFEAIEELIENLWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 343 CGLFEEIEGFIENGWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 344 CGLFEEIEGFIENGWKGLIDWWYGYGHKKHHQHR SEQ ID NO: 345 CFFGAIWEFIKNILKGLIDGWYG SEQ ID NO: 346 CGIFGAIAGFIRSIL SEQ ID NO: 347 CGLFEEIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 348 CGLFEAIEGFIENGWEGMIDGWNGYGRKKRRQRR SEQ ID NO: 349 AGYLLGKINLKALAALAKKILHHHHHHKKKKKKC SEQ ID NO: 350 Bis CGLFEAIEGFIENGWEGMIDWWYGYGRKKRRQRR SEQ ID NO: 351 CGLFEAIEGFIENGWEGMIDGWWYG-(PEG)6-YGRKKRRQRR SEQ ID NO: 352 CGIFGAIWNGIKSLFEGLIDGWYGYGRKKRRQRR SEQ ID NO: 353 CGIFGAIEGFIENGWEGLIDWWYGYGRKKRRQRR SEQ ID NO: 354 CIFGAIAGFIKNIWEGLIDWWYGYGRKKRRQRR SEQ ID NO: 355 CGLFEAIEGFIENGWKGLIDGWWYGGLFEAIEGFIENGWKGLIDWWYG SEQ ID NO: 356 CWEAALAEALAEALAEHLAEALAEALEALAAYGRKKRRQRRK (stearyl) SEQ ID NO: 357 CGLFEAIEGFIENGWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 358 CGLFEELEELLEEGWEGLLEAYGRKKRRQRR SEQ ID NO: 359 CGNFEEIEEFIEEGLRNFIDWWYGYGHKKHHQHR SEQ ID NO: 360 CFFGAIWEFIRNILEGLIDWWYGYGRKKRRQRR SEQ ID NO: 361 CFFGAIWEFIKNILLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 362 CGLFEAIEGFIENGWEGMIDGWWYGYGRKKRRQRR all(D) SEQ ID NO: 363 CGFFHAFFHFFHSFWHGFFEA SEQ ID NO: 364 CGLFHALLHLLHSLWHGLLHWWYGYGHKKHHQHR SEQ ID NO: 365 CGLFGALLELLESLWEGLLEWYGRKKRRQRR SEQ ID NO: 366 CGLFGALLELLESLWEGLLEWYGHKKHHQHR SEQ ID NO: 367 CGLFHALLHLLHSLWKGLLEWWYGF SEQ ID NO: 368 CIFGAIAGFIRSILEGF SEQ ID NO: 369 CGIFGAIAGFIKNIWKGLIDA SEQ ID NO: 370 CFFEAIEEFIKNIWK SEQ ID NO: 371 CGLFEAIEGFIENGWKGLIDWLAEALAEALEALAA SEQ ID NO: 372 GCGIFGAIAEFIKNIW SEQ ID NO: 373 CIFGAIAEFIKNIWKGLIDW SEQ ID NO: 374 CFFGAIWEFIKSILELLLEAYGHKKHHQHRR SEQ ID NO: 375 CWFGAIWEFIKSIL SEQ ID NO: 376 CAFGAIWEFIKSIL SEQ ID NO: 377 CFLGAIWEFIKSIL SEQ ID NO: 378 CFFGAIWEFIKSIK SEQ ID NO: 379 CGFIGAIANLLSKIFEGLIDGWWYGYGRKKRRQRR all(D) SEQ ID NO: 380 CFFGAIWEFIKSIL SEQ ID NO: 381 CIFGAIAGFIKNIWLHLLHHLLHHLHHLLHHLLHL all(D) SEQ ID NO: 382 CFFGAIAEFIKNIW SEQ ID NO: 383 CIFEAIWGFIKNIW SEQ ID NO: 384 stearyl-AGYLLGKINLKALAALAKKILHHHHHHKKKKKKC SEQ ID NO: 385 CIFEAIAGFIKNIWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 386 CGLFEAIEGFIENGWKGLIDWWYGGRPRESGKKRKRKRLKP SEQ ID NO: 387 C(b-Ala)GFGEIEEFIENGLKNLIDWWYGYGHKKHHQHR SEQ ID NO: 388 C(b-Ala)GFEFIEEFIENGLKNLIDWWYGYGRKKRRQRR SEQ ID NO: 389 C(b-Ala)GFEFIEEFIENGLKNLIDWWYGYGHKKHHQHR SEQ ID NO: 390 CGGIEEIAGLLSKILKGLIDWWYGYGHKKHHQHR SEQ ID NO: 391 CGFIGAIANLLSKIFEGLIDWWYGYGRKKRRQRR SEQ ID NO: 392 CGFIGAIAELLEKIFEGLIDWWYGYGRKKRRQRR SEQ ID NO: 393 CGFIGAIAELLEKIFEGLIDWWYGYGHKKHHQHR SEQ ID NO: 394 CFFGAIWEFIRNILEGLIDWWYGYGHKKHHQHR SEQ ID NO: 395 CFFGAIWEFIKSILLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 396 CFFGAIWEFIRSILLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 397 CGFFGAIWEFIRSILEGFIDWWYGYGYGHKKHHQHR SEQ ID NO: 398 CGLFEAIWEFIKSILEGLLEAYGHKKHHQHR SEQ ID NO: 399 CGLFEAIWEFIKSILEGLLEAWYGYGHKKHHQHR SEQ ID NO: 400 CGIFGAIAGFIKNIWKYGRKKRRQRR SEQ ID NO: 401 CGLFEALLELLESLWELLLEAWWYGYGHKKHHQHR SEQ ID NO: 402 CIFGAIAGFIRNIWKGLIDGWYG SEQ ID NO: 403 CGIFGAIAGFIRNIWKGLIDGWYG SEQ ID NO: 404 CFFGAIWEFIKNILKLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 405 CFFGAIWEFIRNILLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 406 CFFGKIWEFIKSIL SEQ ID NO: 407 CYGRKKRRQRRGLFEALLELLESLWELLLEA SEQ ID NO: 408 FFGAIWEFIKSILC SEQ ID NO: 409 CWWGAIEGFIKSIL SEQ ID NO: 410 CFFGAIWEWIKSIL SEQ ID NO: 411 CFFGAIWEFWKSIL SEQ ID NO: 412 CFFGAIWEFIKFIL SEQ ID NO: 413 CFFGAIWEFIKKIL SEQ ID NO: 414 CFFGAIWEFIKGIL SEQ ID NO: 415 CFFGAIWEFIKLIL SEQ ID NO: 416 CFFGAIWEFIKWIL SEQ ID NO: 417 CFFGAIWEFIKSFL SEQ ID NO: 418 CFFGAIWEFIKSKL SEQ ID NO: 419 CFFGFIWEFIKSIL SEQ ID NO: 420 CIFGAIAGFIKNILKGLIDAF SEQ ID NO: 421 CFFGKIWELWEWIL SEQ ID NO: 422 CFFGAIWEFAKSIL SEQ ID NO: 423 CFFGAIWEFIKSAL SEQ ID NO: 424 CFFGAIWEFIKSWL SEQ ID NO: 425 CFFGAIWEFIKSILK SEQ ID NO: 426 CFFGAIWEFIKSILE SEQ ID NO: 427 CFFKAIWEFIKSIL SEQ ID NO: 428 CFFNAIWEFIKSIL SEQ ID NO: 429 CFFGGIWEFIKSIL SEQ ID NO: 430 CFFGNIWEFIKSIL SEQ ID NO: 431 CFFGALWEFIKSIL SEQ ID NO: 432 CFFGAAWEFIKSIL SEQ ID NO: 433 CGLFHALLHLLHSLWHGLLDG SEQ ID NO: 434 CGLFHALLHLLHSLWHGLLEW SEQ ID NO: 435 CGLFHALLHLLHSLWHLLLEA SEQ ID NO: 436 CGLFHALLHLLHSLWKLLLEW SEQ ID NO: 437 CKFGAIWEFIKSIL SEQ ID NO: 438 CFKGAIWEFIKSIL SEQ ID NO: 439 CFFGAIWKFIKSIL SEQ ID NO: 440 CFFGAIWAFIKSIL SEQ ID NO: 441 CFFGAIWLFIKSIL SEQ ID NO: 442 CFFGAIWFFIKSIL SEQ ID NO: 443 CFFGAIWNFIKSIL SEQ ID NO: 444 CFFGAIWELIKSIL SEQ ID NO: 445 CFFGAIWEAIKSIL SEQ ID NO: 446 CGLFEAIEGFIENGWEGLAEALAEALEALAAYGRKKRRQRR SEQ ID NO: 447 CIFGAIAGFIKNIWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 448 CIFGAIAGFIKNIWEGLIDAWYGYGRKKRRQRR SEQ ID NO: 449 CIFGAIAGFIKNIWKGLIDAWYGYGRKKRRQRR SEQ ID NO: 450 CIFGAIAGFIKNIWIFGAIAGFIKNIWWYGYGRKKRRQRR SEQ ID NO: 451 CGLFGAIAGFIENGWEGLIEGWYG SEQ ID NO: 452 CGLFEAIEGFIENGWEGLIDGWYGYGOOOOOQRR (O = ornithine) SEQ ID NO: 453 CGLFEAIEGFIENGWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 454 CGLFEAIEGFIENGWEGLIDGWWYGYGRKKRRQRRK(stearyl) SEQ ID NO: 455 CYGHKKHHQHRGLFEAIEGFIENGWKGLIDWWYG SEQ ID NO: 456 CYGHKKHHQHRGLFEAIEEFIENGWEGLIDGWYG SEQ ID NO: 457 CGLFEAIEGFIENGWKGLIDGWWYGYGRKKRRQRRK(stearyl) SEQ ID NO: 458 CGLFEAIEGFIENGWHGMIDGWWYGYGRKKRRQRR SEQ ID NO: 459 IFGIDDLIIGLLFVAIVEAGIGGYLLGSYGRKKRRQRRC SEQ ID NO: 460 CGFFGEIAELIEEGLKGLIDWWNG SEQ ID NO: 461 CGLFGEIEELIEEGLENLIDWWNG SEQ ID NO: 462 CFFGAIWEFIHSIL all (D) SEQ ID NO: 463 CFFGAIWEFIHNIL SEQ ID NO: 4.64 CFFGAIWEFIHSIFK SEQ ID NO: 4.65 CGIFEAIAGLLKWIFK SEQ ID NO: 466 CGIFELIAGLLKNIFK SEQ ID NO: 467 CGIFEAIAGLLKSILKK (stearyl) SEQ ID NO: 468 CGIFGAIAGLLKSILKK (stearyl) SEQ ID NO: 469 CIFGAIAGFIKNILKGL all (D) SEQ ID NO: 470 CIFGAIAGFIKNILKGLIDGWWYG SEQ ID NO: 471 CIFGAIAGFIKNIWHGLI SEQ ID NO: 472 CIFGAIAGFIKNILKGLK (stearyl) SEQ ID NO: 473 GLGKLINKIFGAIAGFIC all (D) SEQ ID NO: 474 CGIFEAIAGLLKNIFD SEQ ID NO: 475 CGIFEAIAGLLKNIFE SEQ ID NO: 476 CGIFEAIAGLLKNIFR SEQ ID NO: 477 CGIFEAIAGLLKNIFH SEQ ID NO: 478 CGIFEAIAGLLKNIFO (O = ORNITHINE) SEQ ID NO: 479 CGIFEAIAGLLKNIFN SEQ ID NO: 480 CGIFEAIAGLLKNIFCit (Cit = citrulline) SEQ ID NO: 481 CGIFEAIWGLLKNIFK SEQ ID NO: 482 CGIFGAIWGLLKNIFK SEQ ID NO: 4.83 CIFGAIAGLLKNIFK SEQ ID NO: 484 CIFEAIAGLLKNIFK SEQ ID NO: 485 CFFGAIAGLLKNIFK SEQ ID NO: 486 CFFEAIAGLLKNIFK SEQ ID NO: 487 CGFFEAIAGLLKNIFK SEQ ID NO: 488 CIFGAIAGFIKNIWEGLI all (D) SEQ ID NO: 489 CIFGAIAGLLKNIFK all(D) SEQ ID NO: 490 CGLFGEIEELIEEGLENLIDWWNG all(D) SEQ ID NO: 491 CGNFGEIEELIEEGLENLIDWWNG all(D) SEQ ID NO: 492 CGFFGEIAELIEEGLKGLIDWWNG all(D) SEQ ID NO: 493 CGLFGEIEELIEEGLENLIDWWWNE SEQ ID NO: 494 CGFFGAIAGLLKNIFK SEQ ID NO: 495 CGLFELIEGFIENGWEGMIDGWYGYGRKKRRQRRK (stearyl) SEQ ID NO: 496 CGLFELIEGFIEWGWEGMIDGWYGYGRKKRRQRRK (stearyl) SEQ ID NO: 497 CGLFEAIEGFIENGWEGMIDGWWYGYGRKKRRQRRK (2H, 2H, 3H, 3H- SEQ ID NO: 498 perfluorononanoyl) CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRK (2H, 2H, 3H, 3H-perfluoro-10 SEQ ID NO: 499 methylundecanoyl) CIFGAIAGFIKNIWEGLIK (2H, 2H, 3H, 3H-perfluorononanoyl) SEQ ID NO: 500 CIFGAIAGFIKNIWEGLIK (2H, 2H, 3H, 3H-perfluoro-10 methylundecanoyl) SEQ ID NO: 501 CGLFEAIEGFIEWGWEGMIDGWYGYGRKKRRQRRK (2H, 2H, 3H, 3H- SEQ ID NO: 502 perfluorononanoyl) CGLFEAIEGFIEWGWEGMIDGWWYGYGRKKRRQRRK (2H, 2H, 3H,3 H-perfluoro-10 SEQ ID NO: 503 methylundecanoyl) CGLFELIEGFIENGWEGMIDGWWYGYGRKKRRQRRK (2H, 2H, 3H, 3H- SEQ ID NO: 504 perfluorononanoyl) CGLFELIEGFIENGWEGMIDGWYGYGRKKRRQRRK (2H, 2H, 3H, 3H-perfluoro-10 SEQ ID NO: 505 methylundecanoyl) CFFGAIWEFIHSILK (2H, 2H, 3H, 3H-perfluorononanoyl) SEQ ID NO: 506 CFFGAIWEFIHSILK (2H, 2H, 3H, 3H-perfluoro-10 methylundecanoyl) SEQ ID NO: 507 CIFGAIAGFIKNILKGLK (2H, 2H, 3H, 3H-perfluorononanoyl) SEQ ID NO: 508 CIFGAIAGFIKNILKGLK (2H, 2H, 3H, 3H-perfluoro-10 methylundecanoyl) SEQ ID NO: 509 CFFGAIWEFIRNILEGFK (2H, 2H, 3H, 3H-perfluorononanoyl) SEQ ID NO: 510 CFFGAIWEFIRNILEGFK (2H, 2H, 3H, 3H-perfluoro-10 methylundecanoyl) SEQ ID NO: 511 CGLFGEIEELIEEGLENLIDWWNQ SEQ ID NO: 512 CGIFGAIAGLLKSALK SEQ ID NO: 513 CGIFEAIAGLLKSIWK SEQ ID NO: 514 CGIFEAIAGLLKSILK SEQ ID NO: 515 CGIFEAIAGLLONIFK (O = Ornithine) SEQ ID NO: 516 CGIFEAIAGLLKNILKGLIDGWWYG SEQ ID NO: 517 CGIFGAIAGLLKNILKGLIDGWWYG SEQ ID NO: 518 CGIFGAIAGLLKNIFKGLIDGWWYG SEQ ID NO: 519 CGIFGAIWELWEWILK SEQ ID NO: 520 CGIFEAIWELWEWILK SEQ ID NO: 521 CGLFEAIEGFIENGWEGMIDGWYGK (stearyl) SEQ ID NO: 522 (stearyl)GLFEAIEGFIENGWEGMIDGWYGC SEQ ID NO: 523 CFLE-Aib-LWKLLEHLL SEQ ID NO: 524 CFLE-Aib-LWELLEHLL SEQ ID NO: 525 CFLEALWE-Aib-LEHLL SEQ ID NO: 526 CFLE-Aib-LWE-Aib-LEHLL SEQ ID NO: 527 CFLE-Aib-LWEALEKLF SEQ ID NO: 528 (stearyI)IFGAIAGFIKNIWEGLIC SEQ ID NO: 529 CIFGAIAGFIKNIWEGLIK(stearyl) SEQ ID NO: 530 (stearyl)FFGAIWEFIKSILC SEQ ID NO: 531 CFFGAIWEFIKSILK(stearyl) SEQ ID NO: 532 (stearyl)FFGAIWEFIHSILC SEQ ID NO: 533 CFFGAIWEFIHSILK(stearyl) SEQ ID NO: 534 (stearyl)GIFEAIAGLLKNIFKC SEQ ID NO: 535 CGIFEAIAGLLKNIFK(stearyl) SEQ ID NO: 536 CGIFEAIAGLLKNIFKK(stearyl) SEQ ID NO: 537 (stearyl)IFGAIAGFIKNILKGLC SEQ ID NO: 538 CIFGAIAGFIKNILKGLK(stearyl) SEQ ID NO: 539 CIFGAIAGFIKNILKGL SEQ ID NO: 540 CGLFGEIEELIEEGLENLIDWWNS SEQ ID NO: 541 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 542 CGFFGEIAELIEEGLKNLIDWWNG SEQ ID NO: 543 CGLFEAIEGFIENGWKGMIDGWYGYGRKKRRQRR SEQ ID NO: 544 CGLFEAIEGFIEWGWEGMIDGWWYGYGRKKRRQRR SEQ ID NO: 545 CGLFELIEGFIENGWEGMIDGWWYGYGRKKRRQRR SEQ ID NO: 546 CIFGAIAGFIKNIWEGLI SEQ ID NO: 547 CGLFGEIEELIEEGLENLIDWWNG SEQ ID NO: 548 CGLFEEIEGFIENGWEGLIDWWYGYGHKKGGQHR SEQ ID NO: 549 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRK(stearyl) SEQ ID NO: 550 CGLFEALLELLESLWELLEAYGRKKRRQRR SEQ ID NO: 551 CGLFEALLELLESLWELLEAYGRKKRRQRR SEQ ID NO: 552 CFFGAIWEFIRNILEGF SEQ ID NO: 553 CFFGAIWEFIRNILEGFK(stearyl) SEQ ID NO: 554 CIFGAIAGFKNIWEGLIK(lauryl) SEQ ID NO: 555 (lauryl)FFGAIWEFIKSILC SEQ ID NO: 556 CFFGAIWEFIKSILK(lauryl) SEQ ID NO: 557 (lauryl)FFGAIWEFIHSILC SEQ ID NO: 558 CFFGAIWEFIHSILK(lauryl) SEQ ID NO: 559 (lauryl)GIFEAIAGLLKNIFKC SEQ ID NO: 560 CGIFEAIAGLLKNIFK(lauryl) SEQ ID NO: 561 CFFGAIWEFIRNILEGFK(lauryl) SEQ ID NO: 562 (lauryl)GLFEAIEGFIENGWEGMIDGWWYGC SEQ ID NO: 563 CGLFEAIEGFIENGWEGMIDGWYGK(lauryl) SEQ ID NO: 564 CGKFTIVFPHNQKGNWKNVPSNYHYK(stearyl) SEQ ID NO: 565 CMDGTLFPGDDDLAIPATEFFSTKAK(stearyl) SEQ ID NO: 566 CNPVENYIDEVLNEVLWWPNINSSNK(stearyl) SEQ ID NO: 567 CVTPHHVLVDEYTGEWVDSQFK(stearyl) SEQ ID NO: 568 CIFGIDDLIIGLLFVAIVEAGIGGYLLGSK(stearyl) SEQ ID NO: 569 CGAAIGLAWIPYFGPAAEK(stearyl) SEQ ID NO: 570 CFAGWWLAGAALGVATAAQITAGIALHK(stearyl) SEQ ID NO: 571 CFLGFLLGVGSAIASGIAVSKVLHLK(stearyl) SEQ ID NO: 572 CFFGAVIGTIALGVATSAQITAGIALAK(stearyl) SEQ ID NO: 573 CFFGAVIGTIALGVATAAQITAGIALAK(stearyl) SEQ ID NO: 574 GLFEAIAGFIENGGWEGMIDGGGK(stearyl) SEQ ID NO: 575 GLFKAIAKFIKGGWKGLIKGWYGK(stearyl) SEQ ID NO: 576 GLFHAIAHFIHGGWHGLIHGWYGK(stearyl) SEQ ID NO: 577 CGLFEAIAEFIENGWEGLIEGWYK(stearyl) SEQ ID NO: 578 CGFFGAIAGFLEGGWEGMIAGWHGK(stearyl) SEQ ID NO: 579 CFAGWWIGLAALGVATAAQVTAAVALVKK(stearyl) SEQ ID NO: 580 CAVGIVGAMFLGFLGAAGSTMGAVSLTLTVQAK(stearyI) SEQ ID NO: 581 CGVFVLGFLGFLATAGSAMGARSLTLSAK(stearyl) SEQ ID NO: 582 CVPFVLGFLGFLGAAGTAMGAAATALTVK(stearyl) SEQ ID NO: 583 CAVPVAVWLVSALAMGAGVAGGITGSMSLASGK(stearyl) SEQ ID NO: 584 CGLASTLTRWAHYNALIRAFK(stearyl) SEQ ID NO: 585 CGPVEDAITAAIGRVADTVGTK(stearyl) SEQ ID NO: 586 CGLGQMLESMIDNTVREVGGAK(stearyl) SEQ ID NO: 587 CGLFEAIEGFIENGWEGMIDGWWYGFK(stearyl) SEQ ID NO: 588 (D)-(cg1)FEAIEGFIENGWEGMIDGWYGYGRKKRR(D)-(qrr) SEQ ID NO: 589 CGODLEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 590 CIFGIDDLIIGLLFVAIVEAGIGGYLLGS(stearyl) SEQ ID NO: 591 CVTVLALGALAGVGVG(stearyl) SEQ ID NO: 592 CLLGRRGWEVLKYWWWWNLLQYWWSQEL(stearyl) SEQ ID NO: 593 CGIFEAIAGLLKNIFD SEQ ID NO: 594 CGIFEAIAGLLKNIFE SEQ ID NO: 595 CGIFEAIAGLLKNIFR SEQ ID NO: 596 CGIFEAIAGLLKNIFH SEQ ID NO: 597 CGIFEAIAGLLKNIFO (O = ORNITHINE) SEQ ID NO: 598 CGIFEAIAGLLKNIFN SEQ ID NO: 599 CGIFEAIAGLLKNIFCit (Cit = citrulline) SEQ ID NO: 600 CGIFGAIWGLLKNIFK SEQ ID NO: 601 CIFEAIAGLLKNIFK SEQ ID NO: 602 CFFEAIAGLLKNIFK SEQ ID NO: 603 CGFFEAIAGLLKNIFK SEQ ID NO: 604 CGIFEAIAGLLKNIFKG SEQ ID NO: 605 CGIFEAIAGLLKNIFKGL SEQ ID NO: 606 CGIFEAIAGLLKNIFKGLI SEQ ID NO: 607 CGIFEAIAGLLKNIFKGLID SEQ ID NO: 608 CGIFEAIAGLLKNIFKGLIDG SEQ ID NO: 609 CGIFEAIAGLLKNIFKGLIDGF SEQ ID NO: 610 CGIFEAIAGLLKNIFKGLIDGWWYG SEQ ID NO: 611 CGIFEAIAGLLKNIFK SEQ ID NO:612 CGIFEAIAGLLKSILK SEQ ID NO:613 CGIFEAIAGLLKNIFKA SEQ ID NO:614 CGIFEAIAGLLKNIFKL SEQ ID NO:615 CGIFEAIAGLLKNIFKW SEQ ID NO:616 CGIFEAIAGLLKNIFKF SEQ ID NO:617 CGIFEAIAGLLKNAFK SEQ ID NO: 618 CGIFGAIAGLLKNAFK SEQ ID NO: 619 CGIFEAIAGLLONIFO (O = Ornithine) SEQ ID NO: 620 CGIFEAIAGLLKNIFKGIFEAIAGLLKNIFK SEQ ID NO: 621 CGIFEAIAGLLKNIFKFFGAIWEFIHSIL SEQ ID NO: 622 CFFGAIWEFIHSILGIFEAIAGLLKNIFK SEQ ID NO: 623 CFFGAIWEFIHSILFFGAIWEFIHSIL SEQ ID NO: 624 CFFGAIWEFIHSILGFFGAIWEFIHSIL SEQ ID NO: 625 CGIFEAIAGLLKNIFKGIFEAIAGLLKNIFK SEQ ID NO: 626 CGIFEAIAGLLKNIFKFFGAIWEFIHSIL SEQ ID NO: 627 CFFGAIWEFIHSILGIFEAIAGLLKNIFK SEQ ID NO: 628 CGLFHALLHLLHSLWHLLLEA SEQ ID NO: 629 CGLFHALLHLLHSLWHLLLEAK(stearyl) SEQ ID NO: 630 CGLFHALLHLLHSLWHLLLEAK(stearyl) SEQ ID NO: 631 (stearyl)GLFHALLHLLHSLWHLLLEAC SEQ ID NO: 632 CFFGNIWEFIKSIL SEQ ID NO: 633 CFFGAIWLFIKSIL SEQ ID NO: 634 CFFGAIWNFIKSIL SEQ ID NO: 635 CFFGAIWGFIKSIL SEQ ID NO: 636 CFLGALFKALSKLL SEQ ID NO: 637 CFLGALFHALSKLL SEQ ID NO: 638 CFLGALFKALSHLL SEQ ID NO: 639 CFLGALFHALSHLL SEQ ID NO: 640 FLGALFKALSKLLC SEQ ID NO: 641 FLGALFHALSKLLC SEQ ID NO: 642 FLGALFKALSHLLC SEQ ID NO: 643 FLGALFHALSHLLC SEQ ID NO: 644 CFLGALFKALKSLL SEQ ID NO: 645 CFLGALFHALKSLL SEQ ID NO: 646 CFLGALFKALHSLL SEQ ID NO: 647 CFLGALFHALHSLL SEQ ID NO: 648 FLGALFKALKSLLC SEQ ID NO: 649 FLGALFHALKSLLC SEQ ID NO: 650 FLGALFKALHSLLC SEQ ID NO: 651 FLGALFHALHSLLC SEQ ID NO: 652 CGIFGAIAGFIKNIWKGLIDW SEQ ID NO: 653 CGLFEAIEGFIENGWEG-Nle-IDGWYGYGRKKRRQRR SEQ ID NO: 654 CGLFEAIEGFIENGLKGLIDWWYGYGRKKRRQRR SEQ ID NO: 655 CGLFEAIEGFIENAWEGMIDGWWYGYGRKKRRQRR SEQ ID NO: 656 CGLFEAIEGFIENGWEGMIDLWYGYGRKKRRQRR SEQ ID NO: 657 CRLLRLLLRLWRRLLRLLR SEQ ID NO: 658 CGIFGAIEGFIENGWKGLIDAWYGYRKKRRQRR SEQ ID NO: 659 CFFGAIWEFAHGIL SEQ ID NO: 660 CFFGAIWEFARGILEGF SEQ ID NO: 661 FFGAIWEFAHGILC SEQ ID NO: 662 FFGAIWEFARGILEGFC SEQ ID NO: 663 CFFGAIWEFAHSIL SEQ ID NO: 664 FFGAIWEFAHSILC SEQ ID NO: 665 CFFGAIWEFARSILK SEQ ID NO: 666 FFGAIWEFARSILKC SEQ ID NO: 667 CGIFEAIAGLAKNIFK SEQ ID NO: 668 GIFEAIAGLAKNIFKC SEQ ID NO: 669 CGIFEAIAGLAKNIFH SEQ ID NO: 670 CGIFEAIAGLAHNIFH SEQ ID NO: 671 CGIFEAIAGLAHNIFK SEQ ID NO: 672 GIFEAIAGLAKNIFHC SEQ ID NO: 673 GIFEAIAGLAHNIFHC SEQ ID NO: 674 CFLGALWKALSKLL SEQ ID NO: 675 CFLGALWHALSKLL SEQ ID NO: 676 CFLGALWKALSHLL SEQ ID NO: 677 CFLGALWHALSHLL SEQ ID NO: 678 FLGALWKALSKLLC SEQ ID NO: 679 FLGALWHALSKLLC SEQ ID NO: 680 FLGALWKALSHLLC SEQ ID NO: 681 FLGALWHALSHLLC SEQ ID NO: 682 CGIFGAIAGLLKNAFK SEQ ID NO: 683 CIFEAIAGLLKNAFK SEQ ID NO: 684 CIFGAIAGLLKNAFK SEQ ID NO: 685 CIFEAIWEFIKNIW SEQ ID NO: 686 CIFEAIAEFIKNIW SEQ ID NO: 687 CIFGAIWEFIKNIW SEQ ID NO: 688 CIFGAIAEFIKNIW SEQ ID NO: 689 CGIFGIAIGFKINIW SEQ ID NO: 690 CGIFEAIAGLLHNIFK SEQ ID NO: 691 CGIFEAIWGLLHNIFK SEQ ID NO: 692 CGFFEAIAGLLHNIFK SEQ ID NO: 693 CGIFEAIAALLKNIFK SEQ ID NO: 694 CGIFEAIEGLLKNIFK SEQ ID NO: 695 CGIFEAIAGFFKNIFK SEQ ID NO: 696 CGIFEAIAGWWKNIFK SEQ ID NO: 697 CGIFEAIAGLLKNIWK SEQ ID NO: 698 CGIFEAIAELLKNIFK SEQ ID NO: 699 CGIFGAIAGLLKSALK SEQ ID NO: 700 CGIFEAIAGLLKSIWK SEQ ID NO: 701 CGIFEAIAGLLKSILK SEQ ID NO: 702 CGIFEAIAGLLKNIFKGLIDA SEQ ID NO: 703 CGIFEAIAGLLKNIFKGLIDAF SEQ ID NO: 704 CGIFEAIAGLLKNIFKGLIDAWYG SEQ ID NO: 705 CGIFEAIAGLLKNIFKGLIDAWWYGF SEQ ID NO: 706 CGIFEAIAGLLKNIFKGLIDGWYGF SEQ ID NO: 707 CGIFEAIAGLLKNIFKGLIDW SEQ ID NO: 708 CGIFEAIAGLLKNIFKGLIDWF SEQ ID NO: 709 CGIFEAIAGLLKNIFKGLIDWWYG SEQ ID NO: 710 CGIFEAIAGLLKNIFKGLIDWWYGF SEQ ID NO: 711 CGIFELIAGLLKNIFK SEQ ID NO: 712 CGIFEAIAGLLKWIFK SEQ ID NO: 713 CGIFELIAGLLKWIFK SEQ ID NO: 714 CGIFELIAGLLKNIFKG SEQ ID NO: 715 CGIFEAIAGLLKWIFKG SEQ ID NO: 716 CGIFELIAGLLKWIFKG SEQ ID NO: 717 CGLFEALLGLLESLWK SEQ ID NO: 718 CGIFEAIAELLKNIFK SEQ ID NO: 719 CGIFEALLGLLKSLWK SEQ ID NO: 720 CGIFEALLELLKSLWK SEQ ID NO: 721 CGIFEAIAGLLKNIFK SEQ ID NO: 722 CEIFEAIAGLLKNIFK SEQ ID NO: 723 CEIFGAIAGLLKNIFK SEQ ID NO: 724 CGLFEAIAGLLKNLFK SEQ ID NO: 725 CGIWEAIAGLLKNIWK SEQ ID NO: 726 CGLFGAIAGLLKNLFK SEQ ID NO: 727 CGIWGAIAGLLKNIWK SEQ ID NO: 728 CGIFDAIAGLLKNIFK SEQ ID NO: 729 CGIFDAIWGLLKNIFK SEQ ID NO: 730 CGIFGGIGGLLKNIFK SEQ ID NO: 731 CAIFAAIAALLKNIFK SEQ ID NO: 732 CGIFEAIAGLLKNIF SEQ ID NO: 733 CGIFEAIAGLLKNI SEQ ID NO: 734 CGIFEAIAGLLKN SEQ ID NO: 735 CGIFEAIAGLLK SEQ ID NO: 736 CVIFEAIAGLLKNIFK SEQ ID NO: 737 CSIFEAIAGLLKNIFK SEQ ID NO: 738 CGIFEEIAGLLKNIFK SEQ ID NO: 739 CGIFEEIWGLLKNIFK SEQ ID NO: 740 CGIFEAIEELLKNIFK SEQ ID NO: 741 CGIFEAIAGLWKNIFK SEQ ID NO: 742 CGIFEAIAGLLENIFK SEQ ID NO: 743 CGIFEAIAGLLWNIFK SEQ ID NO: 744 CGIFEAIAGLLKEIFK SEQ ID NO: 745 CGIFEAIAGLLKNILK SEQ ID NO: 746 CGIFEAIAGLLRNIFK SEQ ID NO: 747 CGIFEAIAGLLKSIFK SEQ ID NO: 748 CGIFEAIAGLLKNILK SEQ ID NO: 749 CGFFGAIWEFIKSILK SEQ ID NO: 750 CGFFEAIWEFIKSILK SEQ ID NO: 751 CGFFGAIWGLLKSILK SEQ ID NO: 752 CGFFEAIWGLLKSILK SEQ ID NO: 753 CGFFEAIAGLLKSILK SEQ ID NO: 754 CGFFGAIAGLLKSILK SEQ ID NO: 755 CGIFEAIAGLLKNIFEGLI SEQ ID NO: 756 CGIFEAIWGLLKNIFKGLI SEQ ID NO: 757 CGIFEAIWGLLKNIFEGLI SEQ ID NO: 758 CGIFEAIAGLLKNILKGLIDGWWYG SEQ ID NO: 759 CGIFGAIAGLLKNILKGLIDGWWYG SEQ ID NO: 760 CGIFGAIAGLLKNIFKGLIDGWYG SEQ ID NO: 761 CGIFGAIWELWEWILK SEQ ID NO: 762 CGIFEAIWELWEWILK SEQ ID NO: 763 CIFGAIWELWEWILK SEQ ID NO: 764 CIFEAIWELWEWILK SEQ ID NO: 765 CGIFEAIAELWKNIFK SEQ ID NO: 766 CGIFEAIAELWENIFK SEQ ID NO: 767 CGIFEAIAELWKWIFK SEQ ID NO: 768 CGIFEAIAELWEWIFK SEQ ID NO: 769 CGIFEAIAGLLKNILKGLIDWWYG SEQ ID NO: 770 CGIFGAIAGLLKNILKGLIDWWYG SEQ ID NO: 771 CGIFGAIAGLLKNIFKGLIDWWYG SEQ ID NO: 772 CGIFEAIAGLLKNILKGLIDGWYGF SEQ ID NO: 773 CGIFGAIAGLLKNILKGLIDGWYGF SEQ ID NO: 774 CGIFGAIAGLLKNIFKGLIDGWYGF SEQ ID NO: 775 CGIFGAIAELLEKIFE SEQ ID NO: 776 CGIFEAIAELLEKIFE SEQ ID NO: 777 CGFIGAIAELLEKIFE SEQ ID NO: 778 CGIFGAIAELLEKIFK SEQ ID NO: 779 CGIFEAIAELLEKIFK SEQ ID NO: 780 CGFIGAIAELLEKIFK SEQ ID NO: 781 CGLFHALLHLLHSLWHLLLEA SEQ ID NO: 782 GLFHALLHLLHSLWHGLLEAC SEQ ID NO: 783 GFFHAFFHFFHSFWHGFFEAC SEQ ID NO: 784 GLFHALLHLLHSLWHLLLEAC SEQ ID NO: 785 CGLFHALLHLLHSLWHGLLEAK(stearyl) SEQ ID NO: 786 CGFFHAFFHFFHSFWHGFFEAK(stearyl) SEQ ID NO: 787 CGLFHALLHLLHSLWHLLLEAK(stearyl) SEQ ID NO: 788 (stearyl)GLFHALLHLLHSLWHGLLEAC SEQ ID NO: 789 (stearyl)GFFHAFFHFFHSFWHGFFEAC SEQ ID NO: 790 (stearyl)GLFHALLHLLHSLWHLLLEAC SEQ ID NO: 791 CGFFHAFFHFFHSFWHFFFEA SEQ ID NO: 792 CGFFHAFFHFFHSFWHLFFEA SEQ ID NO: 793 CGLFHALLHLLHSLWHGLLEW SEQ ID NO: 794 CGLFHALLHLLHSLWHLLLEW SEQ ID NO: 795 CGFFHAFFHFFHSFWHGFFEW SEQ ID NO: 796 CFFGAIWEFAKSIL SEQ ID NO: 797 CFFGAIWEFAHSIL SEQ ID NO: 798 CFFGAIWEFAHGIL SEQ ID NO: 799 CFFGAIWEFIHSILK SEQ ID NO: 800 CFFGAIWEFIHSILH SEQ ID NO: 801 CFFGAIWEFIHSILD SEQ ID NO: 802 CFFGAIWEFIHSILR SEQ ID NO: 803 CFFGAIWEFIHSILO SEQ ID NO: 804 CFFGAIAEFIHSIL SEQ ID NO: 805 CIFGAIWEFIHSIL SEQ ID NO: 806 CGIFGAIWEFIHSIL SEQ ID NO: 807 CFFGAIWEFIHSILE SEQ ID NO: 808 CFFGAIWEFIHSILEG SEQ ID NO: 809 CFFGAIWEFIHSILEGL SEQ ID NO: 810 CFFGAIWEFIHSILEGLI SEQ ID NO: 811 CFFGAIWEFIHSILEGLID SEQ ID NO: 812 CFFGAIWEFIHSILEGLIDG SEQ ID NO: 813 CFFGAIWEFIHSILEGLIEA SEQ ID NO: 814 CFFGAIWEFIHSILEGLIDW SEQ ID NO: 815 CFFGAIWEFIHSILEGLIDGWYG SEQ ID NO: 816 CFFGAIWEFIHSILEGLIDGWYGF SEQ ID NO: 817 FFGAIWEFIHSILC SEQ ID NO: 818 CFWGAIWEFIHSIL SEQ ID NO: 819 CFFGAIWEFIHSILKGLIDW SEQ ID NO: 820 CAFGKIWEFAHSIL SEQ ID NO: 821 CAFGKIWEFIHSIL SEQ ID NO: 822 CFFGKIWEFIHSIL SEQ ID NO: 823 CAFGAIWEFIHSIL SEQ ID NO: 824 CAFGAIWEFAHSIL SEQ ID NO: 825 CGFFGAIAGLLHNIFK SEQ ID NO: 826 CFFGAIAGLLHNIFK SEQ ID NO: 827 CGFFEAIEGLLHNIFK SEQ ID NO: 828 CFFEAIAGLLHNIFK SEQ ID NO: 829 CFFEAIWGLLHNIFK SEQ ID NO: 830 CGFFGAIAELLHNIFK SEQ ID NO: 831 CFFGAIAELLHNIFK SEQ ID NO: 832 CGFFEAIAELLHNIFK SEQ ID NO: 833 CFFEAIAELLHNIFK SEQ ID NO: 834 CFFGAIWELLHNIFK SEQ ID NO: 835 CFFEAIWELLHNIFK SEQ ID NO: 836 CFFGAIWEFIHSILFFGAIWEFIHSIL SEQ ID NO: 837 CFFGAIWEFIHSILGGGFFGAIWEFIHSIL SEQ ID NO: 838 CFFGAIWEFIHSILGFFGAIWEFIHSIL SEQ ID NO: 839 GGLFEALLELLESLWELLLEW SEQ ID NO: 840 GGFFEAFFEFFESFWEFFFEA SEQ ID NO: 841 GGLFEALLELLESLWEGLLEA SEQ ID NO: 842 CGLFHALLHLLHSLWHLLLHA SEQ ID NO: 843 CGLFEALLHLLHSLWHLLLEA SEQ ID NO: 844 CGLFEALLELLHSLWHLLLEA SEQ ID NO: 845 CGLFEALLHLLESLWHLLLEA SEQ ID NO: 846 CGLFEALLHLLHSLWELLLEA SEQ ID NO: 847 CGLFHALLELLHSLWHLLLEA SEQ ID NO: 848 CGLFHALLHLLESLWHLLLEA SEQ ID NO: 849 CGLFHALLHLLHSLWELLLEA SEQ ID NO: 850 CGLFHALLELLESLWHLLLEA SEQ ID NO: 851 CGLFHALLELLHSLWELLLEA SEQ ID NO: 852 CGLFHALLHLLESLWELLLEA SEQ ID NO: 853 CGLFEALLHLLESLWELLLEA SEQ ID NO: 854 CGLFEALLELLHSLWELLLEA SEQ ID NO: 855 CGLEALLELLESLWHLLLEA SEQ ID NO: 856 CGLFHALLELLESLWELLLEA SEQ ID NO: 857 CFFGAIWEFIHSILHLLLEA SEQ ID NO: 858 CFFGAIWEFIHSILKLLLEA SEQ ID NO: 859 CGFFGAIWEFIHSILGFFGAIWEFIHSIL SEQ ID NO: 860 CFFGAIWEFAHSILFFGAIWEFAHSIL SEQ ID NO: 861 CFFGAIWEFAHSILGFFGAIWEFAHSIL SEQ ID NO: 862 CGFFGAIWEFAHSILGFFGAIWEFAHSIL SEQ ID NO: 863 CFFGAIWEFIHSILGLFEAIEGFIENGWEGMIDG SEQ ID NO: 864 CFFGAIWEFIHSILGLFEAIEGFIENGWEGMIDGWYG SEQ ID NO: 865 CFFGAIWEFIHSILGLFEAIEGFIENGWEGMIDGWWYGF SEQ ID NO: 866 CFFGALLEFIHSILELLLEA SEQ ID NO: 867 CGLFGALLEFIHSILELLLEA SEQ ID NO: 868 CGFFGALLEFIHSILELLLEA SEQ ID NO: 869 CFFGALLEFIHSLWELLLEA SEQ ID NO: 870 CGLFGALLEFIHSLWELLLEA SEQ ID NO: 871 CGFFGALLEFIHSLWELLLEA SEQ ID NO: 872 CIFGAIAGFIKNIWK(stearyl) SEQ ID NO: 873 (stearyl)IFGAIAGFIKNIWC SEQ ID NO: 874 CFFGAIWEFIKSILK(stearyl) SEQ ID NO: 875 (stearyl)FFGAIWEFIKSILC SEQ ID NO: 876 CFFGAIWEFIHSILK(stearyl) SEQ ID NO: 877 (stearyl)FFGAIWEFIHSILC SEQ ID NO: 878 CIFGAIAGFIKNIWEGLIK(stearyl) SEQ ID NO: 879 (stearyl)IFGAIAGFIKNIWEGLIC SEQ ID NO: 880 (stearyl)IFGAIAGFIKNILKGLC SEQ ID NO: 881 (stearyl)GIFGAIAGFIKNILKGLC SEQ ID NO: 882 CIFGAIAGFIKNILKGLK(stearyl) SEQ ID NO: 883 CGLFGAIAGFIVNGWVGMIDG SEQ ID NO: 884 CGLFGAIAGFIVNGWVGMIDGWYG SEQ ID NO: 885 CGLFEAIEGFIVNGWVGMIDGWWYG SEQ ID NO: 886 CGLFGAIAGFIVNGWVGMIDGWWYGF SEQ ID NO: 887 CGLFEAIEAGFIVNGWVGMIDGWYGF SEQ ID NO: 888 CGLFGAIAGFIVNGWVGMIDGWYGK(stearyl) SEQ ID NO: 889 CGLFEAIEGFIVNGWVGMIDGWYGK(stearyl) SEQ ID NO: 890 (stearyl)GLFGAIAGFIVNGWVGMIDGWYGC SEQ ID NO: 891 (stearyl)GLFEAIEGFIVNGWVGMIDGWWYGC SEQ ID NO: 892 (stearyl)GLFGAIAGFIVNGWVGMIDGWYGFC SEQ ID NO: 893 (stearyl)GLFEAIEAGFIVNGWVGMIDGWWYGFC SEQ ID NO: 894 CFFGAIWGLLHSILH SEQ ID NO: 895 CFFGAIWELLHSIL SEQ ID NO: 896 CFFGAIWELLHSILH SEQ ID NO: 897 CFFGAIWGLLHSILK SEQ ID NO: 898 CFFGAIWELLHSILK SEQ ID NO: 899 CGLFGALLHLLHSLWELLLEA SEQ ID NO: 900 CGLFGALLELLHSLWELLLEA SEQ ID NO: 901 CFFGAIWEFIHSILELLLEA SEQ ID NO: 902 CFFGAIWEFIHSILHGLLEA SEQ ID NO: 903 CFFGAIWEFIHSILEGLLEA SEQ ID NO: 904 CGFFGAIWEFIHSILHLLLEA SEQ ID NO: 905 CGFFGAIWEFIHSILELLLEA SEQ ID NO: 906 CGFFGAIWEFIHSILHGLLEA SEQ ID NO: 907 CGFFGAIWEFIHSILEGLLEA SEQ ID NO: 908 CGFFGAIAGLLHSIL SEQ ID NO: 909 CGFFGAIWGLLHSIL SEQ ID NO: 910 CGFFGALLGLLHSIL SEQ ID NO: 911 CFFGAIWEFAKSAL SEQ ID NO: 912 CIFGAIAGFIHNILKGL SEQ ID NO: 913 CFFGAIAGFIKNILKGL SEQ ID NO: 914 CIFGAIWGFIKNILKGL SEQ ID NO: 915 CIFGAIWGFIHNILKGL SEQ ID NO: 916 CIFGAIAGLLKNILKGL SEQ ID NO: 917 CIFGAIAGLLHNILKGL SEQ ID NO: 918 CIFEAIAGFIKNILKGL SEQ ID NO: 919 CIFEAIAGFIHNILKGL SEQ ID NO: 920 CGNFGEIAELIEEGLKNLIDWWNG SEQ ID NO: 921 CGFFGEIAELIEEGLENLIDWWNG SEQ ID NO: 922 CGNFGEIEELIEEGLKNLIDWWNG SEQ ID NO: 923 CGNFGEIAELIEEGLENLIDWWNG SEQ ID NO: 924 CGFFGEIEELIEENGENLIDWWNG SEQ ID NO: 925 CGFFGAIEELIEEGLKNLIDWWNG SEQ ID NO: 926 CGFFGAIAELIEEGLKNLIDWWNG SEQ ID NO: 927 CGFFGEIAELIEEGLKNLIDWWNGF SEQ ID NO: 928 GFFGEIAELIEEGLKNLIDWWNGC SEQ ID NO: 929 GNWWDILNKLGEEILEAIEGFFGC SEQ ID NO: 930 CGNWWDILNKLGEEILEAIEGFFG SEQ ID NO: 931 CGFLGEIAELIEEGLKNLIDWWNG SEQ ID NO: 932 CGFFGEIWELIEEGLKNLIDWWNG SEQ ID NO: 933 CGFFGEIAELWEEGLKNLIDWWNG SEQ ID NO: 934 CGFFGEIAELIWEGLKNLIDWWNG SEQ ID NO: 935 CGFFGEIAELIEWGLKNLIDWWNG SEQ ID NO: 936 CGFFGEIAELIEEGLRNLIDWWNG SEQ ID NO: 937 CGFFGEIAELIEEGLDNLIDWWNG SEQ ID NO: 938 CGFFGEIAELIEEGLKNLNDWWNG SEQ ID NO: 939 CGFFGEIEELIEEGLKNLIDWWNG SEQ ID NO: 940 CGFLGEIEELIEEGLKNLIDWWNG SEQ ID NO: 941 CGFFGLIEELIEEGLKNLIDWWNG SEQ ID NO: 942 CGFFGEIAELIEEGLKNLIDWWNGK(stearyl) SEQ ID NO: 943 (stearyl)GFFGEIAELIEEGLKNLIDWWNGC SEQ ID NO: 944 CFFGAIWEFAKSILK(stearyl) SEQ ID NO: 945 CGFFGAIWEFAKSIL SEQ ID NO: 946 CFFGKIWEFIKSILK(stearyl) SEQ ID NO: 947 (stearyl)FFGKIWEFIKSILC SEQ ID NO: 948 CFFGAIWEFIKSIAK(stearyl) SEQ ID NO: 949 (stearyl)FFGAIWEFIKSIAC SEQ ID NO: 950 (stearyl)FFGAIWEFAKSILC SEQ ID NO: 951 CFFGGIWEFIKSILK(stearyl) SEQ ID NO: 952 (stearyl)FFGGIWEFIKSILC SEQ ID NO: 953 CFFKAIWEFIKSILK(stearyl) SEQ ID NO: 954 (stearyl)FFKAIWEFIKSILC SEQ ID NO: 955 CFFGAIWEAIKSILK(stearyl) SEQ ID NO: 956 (stearyl)FFGAIWEAIKSILC SEQ ID NO: 957 CFFKAIWEFAKSIL SEQ ID NO: 958 CFFKAIWEFAHSIL SEQ ID NO: 959 CFFKAIWEFAKSILK(stearyl) SEQ ID NO: 960 (stearyl)FFKAIWEFAKSILC SEQ ID NO: 961 CFFKAIWEFAHSILK(stearyl) SEQ ID NO: 962 CGLFGEIAELIEEGLENLIDWWNG SEQ ID NO: 963 CGLFGEIEELIEEGLKNLIDWWNG SEQ ID NO: 964 CFFGAIWEFAKSILK(stearyl) SEQ ID NO: 965 CGLFGEIEELIEEGLKGLIDWWNG SEQ ID NO: 966 CGLFGEIAELIEEGLKNLIDWWNG SEQ ID NO: 967 CGLFGEIAELIEEGLEGLIDWWNG SEQ ID NO: 968 GLFGEIEELIEEGLENLIDWWNGC SEQ ID NO: 969 (stearyl)GLFGEIEELIEEGLENLIDWWNGC SEQ ID NO: 970 CGLFGEIEELIEEGLENLIDWWNGK(stearyl) SEQ ID NO: 971 CGMAANDILNELGEEILEEIEGFLG SEQ ID NO: 972 CALFGEIEELIEEGLENLIDWWNG SEQ ID NO: 973 CELFGEIEELIEEGLENLIDWWNG SEQ ID NO: 974 CSLFGEIEELIEEGLENLIDWWNG SEQ ID NO: 975 CNLFGEIEELIEEGLENLIDWWNG SEQ ID NO: 976 CVLFGEIEELIEEGLENLIDWWNG SEQ ID NO: 977 CGFFGEIEELIEEGLENLIDWWNG SEQ ID NO: 978 CGVFGEIEELIEEGLENLIDWWNG SEQ ID NO: 979 CGIFGEIEELIEEGLENLIDWWNG SEQ ID NO: 980 CGWFGEIEELIEEGLENLIDWWNG SEQ ID NO: 981 CGYFGEIEELIEEGLENLIDWWNG SEQ ID NO: 982 CGLLGEIEELIEEGLENLIDWWNG SEQ ID NO: 983 CGLVGEIEELIEEGLENLIDWWNG SEQ ID NO: 984 CGLIGEIEELIEEGLENLIDWWNG SEQ ID NO: 985 CGLWGEIEELIEEGLENLIDWWNG SEQ ID NO: 986 CGLYGEIEELIEEGLENLIDWWNG SEQ ID NO: 987 CGLFEEIEELIEEGLENLIDWWNG SEQ ID NO: 988 CGLFAEIEELIEEGLENLIDWWNG SEQ ID NO: 989 CGLFNEIEELIEEGLENLIDWWNG SEQ ID NO: 990 CGLFSEIEELIEEGLENLIDWWNG SEQ ID NO: 991 CGLFGAIEELIEEGLENLIDWWNG SEQ ID NO: 992 CGLFGDIEELIEEGLENLIDWWNG SEQ ID NO: 993 CGLFGNIEELIEEGLENLIDWWNG SEQ ID NO: 994 CGLFGSIEELIEEGLENLIDWWNG SEQ ID NO: 995 CGLFGELEELIEEGLENLIDWWNG SEQ ID NO: 996 CGLFGEVEELIEEGLENLIDWWNG SEQ ID NO: 997 CGLFGEFEELIEEGLENLIDWWNG SEQ ID NO: 998 CGLFGEWEELIEEGLENLIDWWNG SEQ ID NO: 999 CGLFGEYEELIEEGLENLIDWWNG SEQ ID NO: 1000 CGLFGEIAELIEEGLENLIDWWNG SEQ ID NO: 1001 CGLFGEIGELIEEGLENLIDWWNG SEQ ID NO: 1002 CGLFGEILELIEEGLENLIDWWNG SEQ ID NO: 1003 CGLFGEIVELIEEGLENLIDWWNG SEQ ID NO: 1004 CGLFGEISELIEEGLENLIDWWNG SEQ ID NO: 1005 CGLFGEIEDLIEEGLENLIDWWNG SEQ ID NO: 1006 CGLFGEIENLIEEGLENLIDWWNG SEQ ID NO: 1007 CGLFGEIESLIEEGLENLIDWWNG SEQ ID NO: 1008 CGLFGEIEALIEEGLENLIDWWNG SEQ ID NO: 1009 CGLFGEIEGLIEEGLENLIDWWNG SEQ ID NO: 1010 CGLFGEIEEVIEEGLENLIDWWNG SEQ ID NO: 1011 CGLFGEIEEIIEEGLENLIDWWNG SEQ ID NO: 1012 CGLFGEIEEFIEEGLENLIDWWNG SEQ ID NO: 1013 CGLFGEIEEAIEEGLENLIDWWNG SEQ ID NO: 1014 CGLFGEIEEYIEEGLENLIDWWNG SEQ ID NO: 1015 CGLFGEIEEWIEEGLENLIDWWNG SEQ ID NO: 1016 CGLFGEIEELVEEGLENLIDWWNG SEQ ID NO: 1017 CGLFGEIEELLEEGLENLIDWWNG SEQ ID NO: 1018 CGLFGEIEELFEEGLENLIDWWNG SEQ ID NO: 1019 CGLFGEIEELAEEGLENLIDWWNG SEQ ID NO: 1020 CGLFGEIEELYEEGLENLIDWWNG SEQ ID NO: 1021 CGLFGEIEELWEEGLENLIDWWNG SEQ ID NO: 1022 CGLFGEIEELIDEGLENLIDWWNG SEQ ID NO: 1023 CGLFGEIEELINEGLENLIDWWNG SEQ ID NO: 1024 CGLFGEIEELISEGLENLIDWWNG SEQ ID NO: 1025 CGLFGEIEELIEDGLENLIDWWNG SEQ ID NO: 1026 CGLFGEIEELIEYGLENLIDWWNG SEQ ID NO: 1027 CGLFGEIEELIESGLENLIDWWNG SEQ ID NO: 1028 CGLFGEIEELIEQGLENLIDWWNG SEQ ID NO: 1029 CGLFGEIEELIENGLENLIDWWNG SEQ ID NO: 1030 CGLFGEIEELIEEALENLIDWWNG SEQ ID NO: 1031 CGLFGEIEELIEENLENLIDWWNG SEQ ID NO: 1032 CGLFGEIEELIEESLENLIDWWNG SEQ ID NO: 1033 CGLFGEIEELIEEQLENLIDWWNG SEQ ID NO: 1034 CGLFGEIEELIEEGWENLIDWWNG SEQ ID NO: 1035 CGLFGEIEELIEEGVENLIDWWNG SEQ ID NO: 1036 CGLFGEIEELIEEGIENLIDWWNG SEQ ID NO: 1037 CGLFGEIEELIEEGFENLIDWWNG SEQ ID NO: 1038 CGLFGEIEELIEEGAENLIDWWNG SEQ ID NO: 1039 CGLFGEIEELIEEGYENLIDWWNG SEQ ID NO: 1040 CGLFGEIEELIEEGLRNLIDWWNG SEQ ID NO: 1041 CGLFGEIEELIEEGLHNLIDWWNG SEQ ID NO: 1042 CGLFGEIEELIEEGLONLIDWWNG SEQ ID NO: 1043 CGLFGEIEELIEEGLDNLIDWWNG SEQ ID NO: 1044 CGLFGEIEELIEEGLKNLIDWWNG SEQ ID NO: 1045 CGLFGEIEELIEEGLEGLIDWWNG SEQ ID NO: 1046 CGLFGEIEELIEEGLEYLIDWWNG SEQ ID NO: 1047 CGLFGEIEELIEEGLEQLIDWWNG SEQ ID NO: 1048 CGLFGEIEELIEEGLESLIDWWNG SEQ ID NO: 1049 CGLFGEIEELIEEGLEALIDWWNG SEQ ID NO: 1050 CGLFGEIEELIEEGLE(Cit)LIDWWNG SEQ ID NO: 1051 CGLFGEIEELIEEGLENMIDWWNG SEQ ID NO: 1052 CGLFGEIEELIEEGLENFIDWWNG SEQ ID NO: 1053 CGLFGEIEELIEEGLENIIDWWNG SEQ ID NO: 1054 CGLFGEIEELIEEGLENWIDWWNG SEQ ID NO: 1055 CGLFGEIEELIEEGLENVIDWWNG SEQ ID NO: 1056 CGLFGEIEELIEEGLENYIDWWNG SEQ ID NO: 1057 CGLFGEIEELIEEGLEN(Nle)IDWWNG SEQ ID NO: 1058 CGLFGEIEELIEEGLENLIDWWNG SEQ ID NO: 1059 CGLFGEIEELIEEGLENLVDWWNG SEQ ID NO: 1060 CGLFGEIEELIEEGLENLFDWWNG SEQ ID NO: 1061 CGLFGEIEELIEEGLENLWDWWNG SEQ ID NO: 1062 CGLFGEIEELIEEGLENLYDWWNG SEQ ID NO: 1063 CGLFGEIEELIEEGLENLIEWWNG SEQ ID NO: 1064 CGLFGEIEELIEEGLENLINWWNG SEQ ID NO: 1065 CGLFGEIEELIEEGLENLISWWNG SEQ ID NO: 1066 CGLFGEIEELIEEGLENLIQWWNG SEQ ID NO: 1067 CGLFGEIEELIEEGLENLIDGWNG SEQ ID NO: 1068 CGLFGEIEELIEEGLENLIDAWNG SEQ ID NO: 1069 CGLFGEIEELIEEGLENLIDFWNG SEQ ID NO: 1070 CGLFGEIEELIEEGLENLIDLWNG SEQ ID NO: 1071 CGLFGEIEELIEEGLENLIDIWNG SEQ ID NO: 1072 CGLFGEIEELIEEGLENLIDVWNG SEQ ID NO: 1073 CGLFGEIEELIEEGLENLIDWGNG all (D) SEQ ID NO: 1074 CGLFGEIEELIEEGLENLIDWANG SEQ ID NO: 1075 CGLFGEIEELIEEGLENLIDWFNG SEQ ID NO: 1076 CGLFGEIEELIEEGLENLIDWING SEQ ID NO: 1077 CGLFGEIEELIEEGLENLIDWVNG SEQ ID NO: 1078 CGLFGEIEELIEEGLENLIDWYNG SEQ ID NO: 1079 CGLFGEIEELIEEGLENLIDWWQG SEQ ID NO: 1080 CGLFGEIEELIEEGLENLIDWWTG SEQ ID NO: 1081 CGLFGEIEELIEEGLENLIDWWSG SEQ ID NO: 1082 CGLFGEIEELIEEGLENLIDWWEG SEQ ID NO: 1083 CGLFGEIEELIEEGLENLIDWW(Cit)G SEQ ID NO: 1084 CGLFGEIEELIEEGLENLIDWWNA SEQ ID NO: 1085 CGLFGEIEELIEEGLENLIDWWNN SEQ ID NO: 1086 CGLFGEIEELIEEGLENLIDWWNS SEQ ID NO: 1087 CGLFGEIEELIEEGLENLIDWWNY SEQ ID NO: 1088 CGLFGEIEELIEEGLENLIDWWNW SEQ ID NO: 1089 CFFGAIWGLLHSIL SEQ ID NO: 1090 CFFGK(stearyl)IWEFIKSIL SEQ ID NO: 1091 CFFGK(stearyl)IWEFIHSIL SEQ ID NO: 1092 CFFK(stearyl)AIWEFIKSIL SEQ ID NO: 1093 CGFFGAIWGLLHSILK SEQ ID NO: 1094 CGFFEAIWGLLHSIL SEQ ID NO: 1095 CFFGAIWGLLKSIL SEQ ID NO: 1096 CGFFGAIWGLLKSIL SEQ ID NO: 1097 CFFEAIWGLLKSIL SEQ ID NO: 1098 CGFFEAIWGLLKSIL SEQ ID NO: 1099 CFFGAIWGLLHSILKGLIDWWNG SEQ ID NO: 1100 CFFGAIWGLLHSILKGLIDGWYG SEQ ID NO: 1101 CGIFGAIAGLLKNIFKG SEQ ID NO: 1102 CGIFGAIAGLLKNIFKA SEQ ID NO: 1103 CGIFGAIAGLLKNIFKL SEQ ID NO: 1104 CGIFGAIAGLLKNIFKW SEQ ID NO: 1105 CGIFGAIAGLLKNIFKF SEQ ID NO: 1106 CGIFGAIAGLLKNIFKN SEQ ID NO: 1107 CGIFGAIAGLLKNIFKE SEQ ID NO: 1108 CGIFGAIAGLLKNIFKS SEQ ID NO: 1109 CGIFGAIAGLLKNIFK(stearyl) SEQ ID NO: 1110 CGIFGAIAGLLKNIFKK(stearyl) SEQ ID NO: 1111 (stearyl)GIFGAIAGLLKNIFKC SEQ ID NO: 1112 CGIFGAIAGLLKNIFK(lauryl) SEQ ID NO: 1113 CGIFGAIAGLLKNIFKK(lauryl) SEQ ID NO: 1114 (lauryl)GIFGAIAGLLKNIFKC SEQ ID NO: 1115 CGIFGAIAGLLHNIFK SEQ ID NO: 1116 CGIFGAIAGLLONIFK SEQ ID NO: 1117 CGIFGAIAGLLRNIFK SEQ ID NO: 1118 CGIFGAIAGLLENIFK SEQ ID NO: 1119 CGIFGAIAGLLDNIFK SEQ ID NO: 1120 CGIFGAIAGLLKNIFH SEQ ID NO: 1121 CGIFGAIAGLLKNIFO SEQ ID NO: 1122 CGIFGAIAGLLKINFE SEQ ID NO: 1123 CGIFGAIAGLLKNIFD SEQ ID NO: 1124 CGIFGAIAGLLKNIFN SEQ ID NO: 1125 CGIFGAIAGLLNNIFK SEQ ID NO: 1126 CGIFGIAIGLLKNIFKGIFGAIAGLLKNIFK SEQ ID NO: 1127 CGIFGAIWGLLKNIFKG SEQ ID NO: 1128 CGIFGAIWGLLKNIFKA SEQ ID NO: 1129 CGIFGAIWGLLKNIFKL SEQ ID NO: 1130 CGIFGAIWGLLKNIFKW SEQ ID NO: 1131 CGIFGAIWGLLKNIFKF SEQ ID NO: 1132 CGIFGAIWGLLKNIFKN SEQ ID NO: 1133 CGIFGAIWGLLKNIFKE SEQ ID NO: 1134 CGIFGAIWGLLKNIFKS SEQ ID NO: 1135 CGIFGAIWGLLKNIFK(stearyl) SEQ ID NO: 1136 CGIFGAIWGLLKNIFKK(stearyl) SEQ ID NO: 1137 (stearyl)GIFGAIWGLLKNIFKC SEQ ID NO: 1138 CGIFGAIWGLLKNIFK(lauryl) SEQ ID NO: 1139 CGIFGAIWGLLKNIFKK(lauryl) SEQ ID NO: 1140 (lauryl)GIFGAIWGLLKNIFKC SEQ ID NO: 1141 CGIFGAIWGLLHNIFK SEQ ID NO: 1142 CGIFGAIWGLLONIFK SEQ ID NO: 1143 CGIFGAIWGLLRNIFK SEQ ID NO: 1144 CGIFGAIWGLLENIFK SEQ ID NO: 1145 CGIFGAIWGLLDNIFK SEQ ID NO: 1146 CGIFGAIWGLLKNIFH SEQ ID NO: 1147 CGIFGAIWGLLKNIFO SEQ ID NO: 1148 CGIFGAIWGLLKINFE SEQ ID NO: 1149 CGIFGAIWGLLKNIFD SEQ ID NO: 1150 CGIFGAIWGLLKNIFN SEQ ID NO: 1151 CGIFGAIWGLLNNIFK SEQ ID NO: 1152 CFFGAIWGLLKNIFK SEQ ID NO: 1153 CGFFGAIWGLLKNIFK SEQ ID NO: 1154 CIFGAIWGLLKNIFK SEQ ID NO: 1155 CGIFGAIWIGLLKNIFKGIFGAIWGLLKNIFK SEQ ID NO: 1156 CGIFGAIWGLLHNIFH SEQ ID NO: 1157 CGIFGAIWGLLONIFO SEQ ID NO: 1158 CGIFGAIAGLLHSLK SEQ ID NO: 1159 CGIFGAIWGLLHSILK SEQ ID NO: 1160 CGIFGAIAGLLHSIL SEQ ID NO: 1161 CGIFGAIWGLLHSIL SEQ ID NO: 1162 CGIFGAIWELLKNIFK SEQ ID NO: 1163 CGIFGAIWGLLHNIFHGIFGAIWGLLHNIFK SEQ ID NO: 1164 CGIFEAIWGLLHNIFHGIFEAIWGLLHNIFH SEQ ID NO: 1165 CGIFEAIWGLLKNIFHGIFEAIWGLLHNIFH SEQ ID NO: 1166 CGIFEAIWGLLKNIFKGIFEAIWELLKNIFH SEQ ID NO: 1167 CGIFEAIWGLLKNIFHGIFEAIWGLLKNIFH SEQ ID NO: 1168 CGLFEALLELLESLWELLLEAWNG SEQ ID NO: 1169 CGLFEALLELLESLWELLLEWWNG SEQ ID NO: 1170 CGLFGELEELLEEGLENLLDWWNG SEQ ID NO: 1171 CGLFGELEELLEEGLENLLEWWNG SEQ ID NO: 1172 CGLFGELEELLEEGWELLLEAWNG SEQ ID NO: 1173 CGLFGELEELLEEGWELLLEWWNG SEQ ID NO: 1174 CGLFGELEELLEEGWELLLDWWNG SEQ ID NO: 1175 CGLFGALLELLEEGLENLIDWWNG SEQ ID NO: 1176 CGLFEALLELLEEGLENLIDWWNG SEQ ID NO: 1177 CGLFEALLELLESLLENLIDWWNG SEQ ID NO: 1178 CGLFGELAELLEEGLENLLDWWNG SEQ ID NO: 1179 GLFGEIEELIEEGLENLIDWWNG SEQ ID NO: 1180 CFFGNIWEFIHSIL SEQ ID NO: 1181 CFFGAIWNFIHSIL SEQ ID NO: 1182 CFFGNIWNFIHSIL SEQ ID NO: 1183 CGIFGNIWNFIKNIFK SEQ ID NO: 1184 CGIFGNIWNLLKNIFK SEQ ID NO: 1185 CGIFGNIWGLLKNIFK SEQ ID NO: 1186 CGIFGNIWNFIKNIFH SEQ ID NO: 1187 CGIFGNIWNLLKNIFH SEQ ID NO: 1188 CGIFGNIWGLLKNIFH SEQ ID NO: 1189 CGIFENIWNFIKNIFK SEQ ID NO: 1190 CGIFENIWNFIKNIFH SEQ ID NO: 1191 CGIFENIWGLLKNIFK SEQ ID NO: 1192 CGIFENIWGLLKNIFH SEQ ID NO: 1193 CGIFENIWNLLKNIFK SEQ ID NO: 1194 CGIFENIWNLLKNIFH SEQ ID NO: 1195 CGLFGAIAGLLENIFENLIDWWNG SEQ ID NO: 1196 CGLFGAIAGLLNKIFKNLIDWWNG SEQ ID NO: 1197 CGLFGAIAGLLENIFKNLIDWWNG SEQ ID NO: 1198 CGLFGAIAGLLKNIFENLIDWWNG SEQ ID NO: 1199 CGLFGAIAGLLKNIFHNLIDWWNG SEQ ID NO: 1200 CLIGAILKVLATGLPTLISWIKNKRKQ SEQ ID NO: 1201 CGLLEEIEELLEEGLENLIDWWNG SEQ ID NO: 1202 CGLFEELEELLEEGLENLIDWWNG SEQ ID NO: 1203 CGLFEELEELLEEGLENLIEA SEQ ID NO: 1204 CGLFEELEELLEEGLENLIEAWNG SEQ ID NO: 1205 CGLFEELEELLEEGLENLIEW SEQ ID NO: 1206 CGLFEELEELLEEGLENLIEWWNG SEQ ID NO: 1207 CGLFEELEELLEEGLENLIDA SEQ ID NO: 1208 CGLFEELEELLEEGLENLIDAWNG SEQ ID NO: 1209 CGLFEELEELLEEGLENLIDW SEQ ID NO: 1210 CFLGALKFALKSLL SEQ ID NO: 1211 CFLGALHFALKSLL SEQ ID NO: 1212 CFLGALKFALHSLL SEQ ID NO: 1213 CFLGALHFALHSLL SEQ ID NO: 1214 FLGALKFALKSLLC SEQ ID NO: 1215 GFLGALKFALKSLLC SEQ ID NO: 1216 CGLFGELEELIEEGLENLLDWWNG SEQ ID NO: 1217 CGLFGEIEELLEEGLENLLDWWNG SEQ ID NO: 1218 CGLFGELEELLEEGLENLIDWWNG SEQ ID NO: 1219 CGLFGEIEELIEEGLENLMDWWNG SEQ ID NO: 1220 CGLFGEIEELIEEGLENLEDWWNG SEQ ID NO: 1221 CGLFGEIEELIEEGLENLDDWWNG SEQ ID NO: 1222 CGLFGEIEELIEEGLENLNDWWNG SEQ ID NO: 1223 CGLFGEIEELIEEGLENLSDWWNG SEQ ID NO: 1224 CGLFGEIEELIEEGLENLQDWWNG SEQ ID NO: 1225 CGLFGEIEELIEEGLENL-CIT-DWWNG SEQ ID NO: 1226 CGLFGEIEELIEELLENLIDWWNG SEQ ID NO: 1227 CGLFGEIEELIEEILENLIDWWNG SEQ ID NO: 1228 CGLFGEIEELIEEVLENLIDWWNG SEQ ID NO: 1229 CFLGALWKLLSHLL SEQ ID NO: 1230 CFLGALWKILSHLL SEQ ID NO: 1231 CFLGALWVKVLSHLL SEQ ID NO: 1232 CFLGALWKFLSHLL SEQ ID NO: 1233 CFLEALWKALSHLL SEQ ID NO: 1234 CFLHALWKALSHLL SEQ ID NO: 1235 CFLKALWKALSHLL SEQ ID NO: 1236 CFLNALWKALSHLL SEQ ID NO: 1237 CFLSALWKALSHLL SEQ ID NO: 1238 CFLQALWKALSHLL SEQ ID NO: 1239 CFLEALWEALSHLL SEQ ID NO: 1240 CFLGALWEALSHLL SEQ ID NO: 1241 CFLEALWKLLSHLL SEQ ID NO: 1242 CFLEALWEALEELL SEQ ID NO: 1243 CFLEELWEALEELL SEQ ID NO: 1244 CFLEALWEALEHLL SEQ ID NO: 1245 CFLEELWEALEHLL SEQ ID NO: 1246 CFLEELWELLEELL SEQ ID NO: 1247 CFLEELWELLEHLL SEQ ID NO: 1248 CGLFGEIEELLEEGLE-CIT-LIDWWNG SEQ ID NO: 1249 CGLFEEIEELLEEGLE-CIT-LIDWWNG SEQ ID NO: 1250 CGLFGEIAELLEEGLE-CIT-LIDWWNG SEQ ID NO: 1251 CGLFEEIAELLEEGLE-CIT-LIDWWNG SEQ ID NO: 1252 CGLFGEIEELLEEGLE-CIT-LVDWWNG SEQ ID NO: 1253 CGLFEEIEELLEEGLE-CIT-LVDWWNG SEQ ID NO: 1254 CGLFGEIAELLEEGLE-CIT-LVDWWNG SEQ ID NO: 1255 CGLFEEIAELLEEGLE-CIT-LVDWWNG SEQ ID NO: 1256 CGLFGEIEELLEEGLE-CIT-LIDWWNE SEQ ID NO: 1257 CGLFEEIEELLEEGLE-CIT-LIDWWNE SEQ ID NO: 1258 CGLFGEIAELLEEGLE-CIT-LIDWWNE SEQ ID NO: 1259 CGLFEEIAELLEEGLE-CIT-LIDWWNE SEQ ID NO: 1260 CGLFGEIEELLEEGLH-CIT-LIDWWNG SEQ ID NO: 1261 CGLFEEIEELLEEGLH-CIT-LIDWWNG SEQ ID NO: 1262 CGLFGEIAELLEEGLH-CIT-LIDWWNG SEQ ID NO: 1263 CGLFEEIAELLEEGLH-CIT-LIDWWNG SEQ ID NO: 1264 CGLFGEIEELLEEGLE-CIT-LVDWWNE SEQ ID NO: 1265 CGLFEEIEELLEEGLE-CIT-LVDWWNE SEQ ID NO: 1266 CGLFGEIAELLEEGLE-CIT-LVDWWNE SEQ ID NO: 1267 CGLFEEIAELLEEGLE-CIT-LVDWWNE SEQ ID NO: 1268 CFFKNIWEFIKSIL SEQ ID NO: 1269 CFFKNIWNFIKSIL SEQ ID NO: 1270 CFFKAIWEFIKSILE SEQ ID NO: 1271 CFFKAIWEFIKNIFK SEQ ID NO: 1272 CFFKAIWEFIKNIFKE SEQ ID NO: 1273 CFFKAIWELLKSIL SEQ ID NO: 1274 CFFKAIWGLLKSIL SEQ ID NO: 1275 CFFKAIWEFIKSILK SEQ ID NO: 1276 CFFKNIWGLLKSIL SEQ ID NO: 1277 CFFKAIWGLLKNIFK SEQ ID NO: 1278 CFFKAIWELLKNIFK SEQ ID NO: 1279 CFFKNIWGLLKNIFK SEQ ID NO: 1280 CFFKNIWELLKNIFK SEQ ID NO: 1281 CFFKAIWEFIRSIL SEQ ID NO: 1282 CFFKAIWEFIKSLL SEQ ID NO: 1283 CFFKAIWEFIKSAL SEQ ID NO: 1284 CFFKAIWEFIKSIF SEQ ID NO: 1285 CFFKALWEFLKSLL SEQ ID NO: 1286 CIFKAIWEFIKSIL SEQ ID NO: 1287 CFFKAIWEFIKSIW SEQ ID NO: 1288 CFFHAIWEFIKSIL SEQ ID NO: 1289 CFFEAIWEFIKSIL SEQ ID NO: 1290 CFFKAIAEFIKSIL SEQ ID NO: 1291 CFFKAIEEFIKSIL SEQ ID NO: 1292 CFFKAILEFIKSIL SEQ ID NO: 1293 CFFKAIFEFIKSIL SEQ ID NO: 1294 CFFKAIWGFIKSIL SEQ ID NO: 1295 CFFKAIWHFIKSIL SEQ ID NO: 1296 CFFKAIWKFIKSIL SEQ ID NO: 1297 CFFEAIWKFIKSIL SEQ ID NO: 1298 CFFKAIWELIKSIL SEQ ID NO: 1299 CFFKALWELLKSLL SEQ ID NO: 1300 CFFKAIWEAIKSIL SEQ ID NO: 1301 CFFKAIWEFLKSIL SEQ ID NO: 1302 CFFKAIWEFIHSIL SEQ ID NO: 1303 CFFKAIWEFIESIL SEQ ID NO: 1304 CFFKAIWEFIKNIL SEQ ID NO: 1305 CFFKAIWEFIKWIL SEQ ID NO: 1306 CFFKAIWEFIKEIL SEQ ID NO: 1307 CFFKAIWEFIKGIL SEQ ID NO: 1308 CFFKAIWEFIKSGL SEQ ID NO: 1309 CFFKAIWEFIKSII SEQ ID NO: 1310 CFFKAIWEFIK-CIT-IL SEQ ID NO: 1311 CFFKAIWEFIKSIA SEQ ID NO: 1312 CFFKAIWEFIKQIL SEQ ID NO: 1313 CGFFKAIWEFIKSIL SEQ ID NO: 1314 CFFKAIWEFIKSILKGLIDG SEQ ID NO: 1315 CFFKAIWEFIKSILKGLIDGWYG SEQ ID NO: 1316 CFFKAIWEFIKSILEGLIDG SEQ ID NO: 1317 CFFKAIWEFIKSILEGLIDGWYG SEQ ID NO: 1318 CFFKAIWEFIKNIFKGLIDG SEQ ID NO: 1319 CFFKAIWEFIKNIFKGLIDGWYG SEQ ID NO: 1320 CFFGNIWEFIKSILKGLIDG SEQ ID NO: 1321 CFFGNIWEFIKSILKGLIDGWYG SEQ ID NO: 1322 CFFGNIWEFIKSILEGLIDG SEQ ID NO: 1323 CFFGNIWEFIKSILEGLIDGWYG SEQ ID NO: 1324 CFFGNIWEFIKNIFKGLIDG SEQ ID NO: 1325 CFFGNIWEFIKNIFKGLIDGEYG SEQ ID NO: 1326 CFFKAIWGLLKSILKGLIDG SEQ ID NO: 1327 CFFKAIWGLLKSILKGLIDGWYG SEQ ID NO: 1328 CFFKAIWGLLKSILEGLIDG SEQ ID NO: 1329 CFFKAIWGLLKSILEGLIDGWYG SEQ ID NO: 1330 CFFKAIWGLLKNIFKGLIDG SEQ ID NO: 1331 CFFKAIWGLLKNIFKGLIDGWYG SEQ ID NO: 1332 CFFKAIWGLLKNIFEGLIDG SEQ ID NO: 1333 CFFKAIWGLLKNIFEGLIDGWYG SEQ ID NO: 1334 CFFKAIWEFIKSILKGLIDGWNG SEQ ID NO: 1335 CFFKAIWEFIKNIFKGLIDGWNG SEQ ID NO: 1336 CIFGAIAGLLKNILKGLIDG SEQ ID NO: 1337 CIFGAIAGLLKNILKGLIDGWYG SEQ ID NO: 1338 CFLEALWKALEHLL SEQ ID NO: 1339 CFLEALWEALSKLL SEQ ID NO: 1340 CFLEALWEALEKLL SEQ ID NO: 1341 CFLEALWEALEHLLK(stearyl) SEQ ID NO: 1342 (stearyl)FLEALWEALEHLLC SEQ ID NO: 1343 (stearyl)GFLEALWEALEHLLC SEQ ID NO: 1344 CFLEALWKALSKLL SEQ ID NO: 1345 CFLEALWEALDHLL SEQ ID NO: 1346 CFLEALWEALTHLL SEQ ID NO: 1347 CFLEALWEALNHLL SEQ ID NO: 1348 CFLEALWEALQHLL SEQ ID NO: 1349 CFLEALWEALEHLLH SEQ ID NO: 1350 CFLEALWEALEHLLK SEQ ID NO: 1351 CFLEALWEALEHLLE SEQ ID NO: 1352 CWLEALEALEHLL SEQ ID NO: 1353 CLLEALWEALEHLL SEQ ID NO: 1354 CFFEALWEALEHLL SEQ ID NO: 1355 CFLEALEEALEHLL SEQ ID NO: 1356 CFLEALAEALEHLL SEQ ID NO: 1357 CFLEALFEALEHLL SEQ ID NO: 1358 CLFEALWEALHHLL SEQ ID NO: 1359 CLFEALWEALKHLL SEQ ID NO: 1360 CFLEALWEALEHGL SEQ ID NO: 1361 CLFEALWEALEHLF SEQ ID NO: 1362 CLFEALWEALEHFL SEQ ID NO: 1363 CLFEALWEALEHLLEGLIDWWYG SEQ ID NO: 1364 CLFEALWEALEHLLEGLIDWWNG SEQ ID NO: 1365 CLFEALWEALEHLLENLIDWWNG SEQ ID NO: 1366 CFLEELWELLEKLL SEQ ID NO: 1367 CFLEELWELLEELLE SEQ ID NO: 1368 CFLEELWELLEELLELLE SEQ ID NO: 1369 CFLEELWELLEHLLELLD SEQ ID NO: 1370 CFLEELWELLEELLELID SEQ ID NO: 1371 CFLEELWELLEELLELLD SEQ ID NO: 1372 CFLEELWELLEHLLEGLE SEQ ID NO: 1373 CFLEELWELLEHLLEGLD SEQ ID NO: 1374 CFLEELWELLEHLLEEGLI SEQ ID NO: 1375 CFLEELWELLEHLLEGLIDWWYG SEQ ID NO: 1376 CFLEELWELLEHLLENLIDWWNG SEQ ID NO: 1377 CFLEALWEALEHLLELLD SEQ ID NO: 1378 CGLFGELEELLEEGLENLTDWWNG SEQ ID NO: 1379 CGLFGELEELLEEGLENL-(ALLO-I)-DWWNG SEQ ID NO: 1380 CFLEALWEALEHLLELID SEQ ID NO: 1381 CELFEELEELLEEGLENLIDWWNG SEQ ID NO: 1382 CGLFEELEELLEEGLELLIDWWNG SEQ ID NO: 1383 CGLFEELEELLEEGLELLIDWWNK SEQ ID NO: 1384 CGLFEELEELLEEGLENLIDWWNK SEQ ID NO: 1385 CGLFGELEELLEEGLENLIDWWNQ SEQ ID NO: 1386 CGLFGELEELLEEGLENLIDWWNE SEQ ID NO: 1387 CGLFGELEELLEEGLENLIDWWNN SEQ ID NO: 1388 CGLFGELEELLEEGLENLIDWWNS SEQ ID NO: 1389 CGLFEELEELLEEGLENLIDWWNQ SEQ ID NO: 1390 AC-CFLEELWELLEHLL SEQ ID NO: 1391 AC-CFLEELWELLEELL SEQ ID NO: 1392 CGLLGEIEELLEEGLENLIDWWNG SEQ ID NO: 1393 CGLLAEIEELLEEGLENLIDWWNG SEQ ID NO: 1394 CGLLGEIEELLEEGLENLIDWWNQ SEQ ID NO: 1395 CGLLAEIEELLEEGLENLIDWWNQ SEQ ID NO: 1396 CGLLEEIEELLEEGLENLIDWWNQ SEQ ID NO: 1397 CGLLGEIEELLEEGLENLIDWWNE SEQ ID NO: 1398 CGLLAEIEELLEEGLENLIDWWNE SEQ ID NO: 1399 CGLLEEIEELLEEGLENLIDWWNE SEQ ID NO: 1400 CGLLGEIEELLEEGLENIDWWNS SEQ ID NO: 1401 CGLLAEIEELLEEGLENLIDWWNS SEQ ID NO: 1402 CGLLEEIEELLEEGLENLIDWWNS SEQ ID NO: 1403 CGLFAELEELLEEGLENLLEWWNG SEQ ID NO: 1404 CGLFEELEELLEEGLENLLEWWNG SEQ ID NO: 1405 CGLFGELEELLEEGLENLLEWWNE SEQ ID NO: 1406 CGLFAELEELLEEGLENLLEWWNE SEQ ID NO: 1407 CGLFEELEELLEEGLENLLEWWNE SEQ ID NO: 1408 CGLLGELEELLEEGLENLLEWWNG SEQ ID NO: 1409 CGLLGELEELLEEGLENLLEWWNE SEQ ID NO: 1410 CGILGEIEELLEEGLENLIDWWNG SEQ ID NO: 1411 CGILGEIEELLEEGLENLIDWWNE SEQ ID NO: 1412 CGILGEIEELLEEGLENLIDWWNS SEQ ID NO: 1413 CGILAEIEELLEEGLENLIDWWNG SEQ ID NO: 1414 CGILEEIEELLEEGLENLIDWWNG SEQ ID NO: 1415 CIFGAIAELLKNIFK SEQ ID NO: 1416 CIFGAIAELLENIFK SEQ ID NO: 1417 CIFGAIAGLLENIFK SEQ ID NO: 1418 CFLEELWGLLEHLL SEQ ID NO: 1419 CGILAEIEELLEEGLENLIDWWNQ SEQ ID NO: 1420 CGILAEIEELLEEGLENLIDWWNE SEQ ID NO: 1421 CGLFAEIEELLEEGLENLIDWWNQ SEQ ID NO: 1422 CGLFAEIEELLEEGLENLIDWWNE SEQ ID NO: 1423 CGLFGELEELLEEGLENLLEWWNQ SEQ ID NO: 1424 CGLFAEIAELLEEGLE-CIT-LIDWWNE SEQ ID NO: 1425 CGILAEIEELLEEGLENLLEWWNG SEQ ID NO: 1426 CGILEEIEELLEEGLENLIDWWNE SEQ ID NO: 1427 CGILEEIEELLEEGLENLIDWWNQ SEQ ID NO: 1428 CGLFGEIEELIWEGLENLIDWWNG SEQ ID NO: 1429 CGLFGEIAELIWEGLENLIDWWNG SEQ ID NO: 1430 CGLFEEIAELIEEGLENLIDWWNG SEQ ID NO: 1431 CGLFEEIAELIWEGLENLIDWWNG SEQ ID NO: 1432 CELFEEIAELIWEGLENLIDWWNG SEQ ID NO: 1433 CELFEEIAELLWEGLENLIDWWNG SEQ ID NO: 1434 CGLFEEIAELLWEGLENLIDWWNG SEQ ID NO: 1435 CGLFEELAELLWEGLENLIDWWNG SEQ ID NO: 1436 CELFEELAELLWEGLENLIDWWNG SEQ ID NO: 1437 CELFEELAELLWEGLENLIDWWNS SEQ ID NO: 1438 CGLFEELAELLWEGLENLIDWWNS SEQ ID NO: 1439 CGIFEELAELLWEGLENLIDWWNG SEQ ID NO: 1440 CGIFEELAELLWEGLENLIDWWNS SEQ ID NO: 1441 CGLFEELEELLEELLENLIDWWNS SEQ ID NO: 1442 CELFEELEELLEELLENLIDWWNS SEQ ID NO: 1443 CELFEELEELLEELLELLIDWWNS SEQ ID NO: 1444 CEFLEELEELLEELLENLIDWWNS SEQ ID NO: 1445 CELFEELEELLEHLLENLIDWWNS SEQ ID NO: 1446 CELFEELEELLHELLENLIDWWNS SEQ ID NO: 1447 CGLFGELEELLWEGLENLIDWWNG SEQ ID NO: 1448 CGLFGELEELLWEGLHNLIDWWNG SEQ ID NO: 1449 CGLFGELWELLEHGLENLIDWWNG SEQ ID NO: 1450 CGL-R6H-GELEEL-57H-EEGLENLIDWWNG SEQ ID NO: 1451 CGLFEAIEGFIENGWEGMIDGWNG SEQ ID NO: 1452 CGLFEAIEGFIENGWEGMIDWWNG SEQ ID NO: 1453 CGLFGAIEGFIENGWEGMIDWWNG SEQ ID NO: 1454 CGLFAEIEELLEEGLENLLEWWNG SEQ ID NO: 1455 CGLFAELEELLEEGLENLIDWWNG SEQ ID NO: 1456 CGIFAEIEELLEEGLENLIDWWNG SEQ ID NO: 1457 CGLFAEIEELLEEGLENLIDWWNGF SEQ ID NO: 1458 CGLFAEIEELLEEGLENLIDWWNA SEQ ID NO: 1459 CGLFAEIEELLEEGLENLIDWWNS SEQ ID NO: 1460 CGLFAEIEELLEEGLENLIDWWN-CIT SEQ ID NO: 1461 CGLFGEIAGLLEEGLHNLIDWWNG SEQ ID NO: 1462 CGLFGEIAGLLEQGLHNLIDWWNG SEQ ID NO: 1463 CGLFGEIAGLLESGLHNLIDWWNG SEQ ID NO: 1464 CGLFAEIAGLLEQGLHNLIDWWNG SEQ ID NO: 1465 CGLFAEIAGLLEEGLHNLIDWWNG SEQ ID NO: 1466 CGLFAEIAGLLESGLHNLIDWWNG SEQ ID NO: 1467 CGIFEAIAGLLEQGLHNLIDWWNG SEQ ID NO: 1488 CGLFGAIAELLEEGLHNLIDWWNG SEQ ID NO: 1469 CGLFAAIAELLEEGLHNLIDWWNG SEQ ID NO: 1470 CGIFEAIAGLLKNIFKNLIDWWNG SEQ ID NO: 1471 CGIFGAIWELLEQGLHNLIDWWNG SEQ ID NO: 1472 CGLFAELAGLLEQGLHNLIDWWNG SEQ ID NO: 1473 CGILAELAGLLEQGLHNLIDWWNG SEQ ID NO: 1474 CGLFGEIEELLEHLL SEQ ID NO: 1475 CGLFGEIEELLEELL SEQ ID NO: 1476 CGLFGEIEELLEEGL SEQ ID NO: 1477 CGLFGEIEELLEHGL SEQ ID NO: 1478 CGLFHEIEELLEHLL SEQ ID NO: 1479 CFLGALWKALSELLE SEQ ID NO: 1480 CGLFGEIWELLEEGL SEQ ID NO: 1481 CGLFGEIWELLEEGLI SEQ ID NO: 1482 CGLFGEIWELLEELL SEQ ID NO: 1483 CGLFEEIEELLEELLE SEQ ID NO: 1484 CGLFELIEGFIEWGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 1485 CIFGAIAGFIKNIWLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 1486 CEALFGKINAIFIGKL SEQ ID NO: 1487 CEENWIGLFGGGNIWEEEEILDLL SEQ ID NO: 1488 CLELWLEHLFLELE SEQ ID NO: 1489 CGNFEEIEGFIENGWEGLIDGWWYGYGRKKRRQRR SEQ ID NO: 1490 CRGKWYMGFGEIKRQGEGRRYGLFEDWIAENRGI SEQ ID NO: 1491 GLFEAIEGFIENGWEGLAELAEALEALAAGGSC SEQ ID NO: 1492 GLFGALAEALAEALAEHLAEALAEALEALAAGGSC SEQ ID NO: 1493 CGFFGEIAGLLENGLHNLIDWWNG SEQ ID NO: 1494 CGFFGEIAALLENGLENLIDWWNG SEQ ID NO: 1495 CGFFGEIAEFIHSGLKNLIDWWNG SEQ ID NO: 1496 CGFFGEIAGLLKNGLKNLIDWWNG SEQ ID NO: 1497 CGFFGEIAGFIKNGLKNLIDWWNG SEQ ID NO: 1498 CGFFGEIAEFIHSILKNLIDWWNG SEQ ID NO: 1499 CGFFGEIAGLLKNILKNLIDWWNG SEQ ID NO: 1500 CGFFGEIAGFIKNILKNLIDWWNG SEQ ID NO: 1501 CFLGALFHALSELL SEQ ID NO: 1502 CFLGALWHALSELL SEQ ID NO: 1503 CFLGALWHALSHLL SEQ ID NO: 1504 CFLGALWELLSHLL SEQ ID NO: 1505 CFLGALWKALSHLL SEQ ID NO: 1506 CFLGALWHALSKLL SEQ ID NO: 1507 CFLGALFHLLSHLL SEQ ID NO: 1508 CFLGALFHLLSELL SEQ ID NO: 1509 CFLGALWHLLSHLL SEQ ID NO: 1510 CFLGALWHLLSELL SEQ ID NO: 1511 CFLGALFHALSHLLE SEQ ID NO: 1512 CFLGALFHLLSHLLE SEQ ID NO: 1513 CGLFGALFHALSHLLE SEQ ID NO: 1514 CFLGALWKALSHLL SEQ ID NO: 1515 CGLFAEIEELLEEGLENLIDWWNG SEQ ID NO: 1516 CGLFGEIEELIEEGLE-Cit-LIDWWNG SEQ ID NO: 1517 CGLFGEIEELIEEGLENLIDWWNE SEQ ID NO: 1518 CFFGAIWEFIHSILK(stearyl) SEQ ID NO: 1519 CIFGAIAGFIKNIWEGLIK(stearyl) SEQ ID NO: 1520 CGIFEAIAGLLKNIFK(stearyl) SEQ ID NO: 1521 CGIFEAIAGLLKNIFKK(stearyl) SEQ ID NO: 1522 CFLGALFHALSHLL SEQ ID NO: 1523 Ac-CIFGAIAGFIKNILKGLIDG SEQ ID NO: 1524 CIFGAIAGFIKNILKGLK(stearylL) SEQ ID NO: 1525 Ac-CIFGAIAGFIKNILKGLK(stearyl) SEQ ID NO: 1526 CGLFGEIEELIEEGLENLIDWWNG SEQ ID NO: 1527 CFLGALWKALSELLKNLIDWWNG SEQ ID NO: 1528 CGFLGALWKALSELLKNLIDWWNG SEQ ID NO: 1529 CFLGALFHALSHLLENLIDWWNG SEQ ID NO: 1530 CGFLGALFHALSHLLENLIDWWNG SEQ ID NO: 1531 CGLFGELEGFIENGLKNLIDWWNG SEQ ID NO: 1532 CGLFGELEGLLWHGLKNLIDWWNG SEQ ID NO: 1533 CGLFGELAELLWHGLKNLIDWWNG SEQ ID NO: 1534 CGLFGELAELLWQGLKNLIDWWNG SEQ ID NO: 1535 CGLFGELWELLWHGLKNLIDWWNG SEQ ID NO: 1536 CGLFGELWELLWQGLKNLIDWWNG SEQ ID NO: 1537 CGLFEELAGLLWHGLKNLIDWWNG SEQ ID NO: 1538 CGLFEELWGLLWHGLKNLIDWWNG SEQ ID NO: 1539 CGLFEELAGLLWQGLKNLIDWWNG SEQ ID NO: 1540 CGLFEELWGLLWQGLKNLIDWWNG SEQ ID NO: 1541 CGLFGELAELLWHGLKNLIDWWNK SEQ ID NO: 1542 CGLFEELAELLWHGLKNLIDWWNK SEQ ID NO: 1543 CGLFGELAELLWHGLKNLIDWWNH SEQ ID NO: 1544 CGLFEELAELLWHGLKNLIDWWNH SEQ ID NO: 1545 CGLFAELWGLLWQGLKNLIDWWNG SEQ ID NO: 1546 CGLFAELWGLLWHGLKNLIDWWNG SEQ ID NO: 1547 CGLFAELWGLLWHGLHNLLDWWNG SEQ ID NO: 1548 CGLFAELAELLWEGLKNLIDWWNG SEQ ID NO: 1549 CGLFAELAELLWHGLKNLIDWWNG SEQ ID NO: 1550 CGLFAELELLWQGLKNLIDWWNG SEQ ID NO: 1551 CELFGELAGLLWHGLKNLIDWWNG SEQ ID NO: 1552 CLFEALWE-Aib-LEKLF SEQ ID NO: 1553 CFLEALWELLEHLL SEQ ID NO: 1554 CFLEALWKALEKLL SEQ ID NO: 1555 CGLF-Aib-EIAGLLEEGLHNLIDWWNG SEQ ID NO: 1556 CGLFGEI-Aib-GLLEEGLHNLIDWWNG SEQ ID NO: 1557 CGFFGEIAGLLEE-Aib-LHNLIDWWNG SEQ ID NO: 1558 CGLFGEIAGLLEEGLHNLIDWWN-Aib SEQ ID NO: 1559 CGLF-Aib-EIAGLLEE-Aib-LHNLIDWWNG SEQ ID NO: 1560 CGFFGEI-Aib-GLLEE-Aib-LHNLIDWWNG SEQ ID NO: 1561 CGFFGEI-Aib-ELIWEGLKNLIDWWNG SEQ ID NO: 1562 CGFFGEIAELIWELKNLIDWWN-Aib SEQ ID NO: 1563 CGFFAib-EIAELIWE-Aib-LKNLIDWWNG SEQ ID NO: 1564 AC-CFLGALWKALSHLL SEQ ID NO: 1565 AC-CFLEELWELLEELLE SEQ ID NO: 1566 AC-CLFGALWKALSELL SEQ ID NO: 1567 AC-CGIGAVLKVLTTGLPALISWIKRKRQQ SEQ ID NO: 1568 AC-CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 1569 AC-CGLFEAIEGFIENGWEGMIDGWWYGYGRKKRRQRRK(stearyl) SEQ ID NO: 1570 Ac-CFLGALWKALSHLL SEQ ID NO: 1571 Ac-CFLGALWKALSELL SEQ ID NO: 1572 CELFEEIAELLWEGLENLIDWWNG SEQ ID NO: 1573 CGLFGEIAELIWEGLENLIDWWNG SEQ ID NO: 1574 CGLFGEIEELLEEGLENLIDWWNG SEQ ID NO: 1575 CGLFAELAELLWEGLENLIDWWNG SEQ ID NO: 1576 CGLFAELAELLEEGLENLIDWWNG SEQ ID NO: 1577 CGLFAELAELLWEGLENLIDWWNS SEQ ID NO: 1578 CGLFAELAELLEEGLENLIDWWNS SEQ ID NO: 1579 CGLFAELAELLWEGLENLIDWWNQ SEQ ID NO: 1580 CGLFAELAELLEEGLENLIDWWNQ SEQ ID NO: 1581 CGLFAELAELLWEGLENLIDWWNE SEQ ID NO: 1582 CGLFAELAELLEEGLENLIDWWNE SEQ ID NO: 1583 CELFEELAELLWEGLENLIDWWNQ SEQ ID NO: 1584 CELFEELAELLWEGLENLIDWWNE SEQ ID NO: 1585 CELFEELAELLEEGLENLIDWWNG SEQ ID NO: 1586 CELFAELAELLWEGLENLIDWWNG SEQ ID NO: 1587 CELFAELAELLEEGLENLIDWWNG SEQ ID NO: 1588 CELFAELAELLWEGLENLIDWWNS SEQ ID NO: 1589 CELFAELAELLEEGLENLIDWWNS SEQ ID NO: 1590 CELFAELAELLWEGLENLIDWWNQ SEQ ID NO: 1591 CELFAELAELLEEGLENLIDWWNQ SEQ ID NO: 1592 CELFAELAELLWEGLENLIDWWNE SEQ ID NO: 1593 CELFAELAELLEEGLENLIDWWNE SEQ ID NO: 1594 CELFEELAELLWEGLHNLIDWWNG SEQ ID NO: 1595 CELFEELAELLWEGLHNLIDWWNS SEQ ID NO: 1596 CELFEELAELLWEGLHNLIDWWNQ SEQ ID NO: 1597 CELFEELAELLWEGLHNLIDWWNE SEQ ID NO: 1598 CELFGELEGFIENGLENLIDWWNG SEQ ID NO: 1599 CGLFEELEGFIENGLENLIDWWNG SEQ ID NO: 1600 CGLFAELAGFIENGLENLIDWWNG SEQ ID NO: 1601 CGLFAELEGFIENGLENLIDWWNG SEQ ID NO: 1602 CGLFGELAGFIENGLENLIDWWNG SEQ ID NO: 1603 CELFEELEGFIENGLENLIDWWNG SEQ ID NO: 1604 CELFAELAGFIENGLENLIDWWNG SEQ ID NO: 1605 CGLFGELEGFIWNGLENLIDWWNG SEQ ID NO: 1606 CGLFGELEGFIENGLENLIDWWNG SEQ ID NO: 1607 CGLFGELEGFIENGLENLIDWWNQ SEQ ID NO: 1608 CGLFGELEGFIENGLENLIDWWNE SEQ ID NO: 1609 CELFEELEGFIENGLENLIDWWNE SEQ ID NO: 1610 CGLLEEIAELLEEGLENLIDWWNS SEQ ID NO: 1611 CGLLEEIEELLWEGLENLIDWWNS SEQ ID NO: 1612 CELLEEIEELLEEGLENLIDWWNS SEQ ID NO: 1613 CGLLEEIAELLWEGLENLIDWWNS SEQ ID NO: 1614 CELLEEIAELLWEGLENLIDWWNS SEQ ID NO: 1615 CELLEEIEELLEEGLENLIDWWNE SEQ ID NO: 1616 CGLLEELEELLEEGLENLIDWWNS SEQ ID NO: 1617 CGLLEELEELLEEGLENLLEWWNS SEQ ID NO: 1618 CGLLEEIAELLEEGLENLIDWWNG SEQ ID NO: 1619 CGLLAEIAELLEEGLENLIDWWNS SEQ ID NO: 1620 CGLLAEIAELLWEGLENLIDWWNS SEQ ID NO: 1621 CGLLEEIEGFIENGLENLIDWWNS SEQ ID NO: 1622 CGLLEEIEGFIENGLENLIDWWNG SEQ ID NO: 1623 CGLLEEIEELLEEGLE-Cit-LIDWWNS SEQ ID NO: 1624 CGLLEEIEELLEQGLENLIDWWNS SEQ ID NO: 1625 CGLLAELAELLEEGLENLIDWWNS SEQ ID NO: 1626 CGLLEEIEELLEEGLENLIDWWNA SEQ ID NO: 1627 CGLL-Aib-EIEELLEEGLENLIDWWNS SEQ ID NO: 1628 CGLLEEIEELLEEGLENLIDWWN-Aib SEQ ID NO: 1629 CGLLEEIEELLEE-Aib-LENLIDWWNG SEQ ID NO: 1630 CGLFGHIHHLIHHGLHNLIDWWNG SEQ ID NO: 1631 CGLFGEIHHLIHHGLHNLIDWWNG SEQ ID NO: 1632 CGLFGEIHHLIHHGLENLIDWWNG SEQ ID NO: 1633 CGLFGEIHELIHHGLENLIDWWNG SEQ ID NO: 1634 CELLEEIEELLEEGLENLIDWWNS SEQ ID NO: 1635 CGLFGELEELIEEGLENLIDWWNG SEQ ID NO: 1636 CGLLAEIEELLWEGLENLIDWWNS SEQ ID NO: 1637 CGLLEEIEELLEEGLENLLEWWNS SEQ ID NO: 1638 C(b-ALA)LLEEIEELLEEGLENLIDWWNS SEQ ID NO: 1639 CGLLEEIEELLEEGLENLIDLWNS SEQ ID NO: 1640 CGLLEEIEELLEWGLENLIDWWNS SEQ ID NO: 1641 CGLFGEIEELIEEGLENLIDWGNG SEQ ID NO: 1642 CGFFGEIAELIEEGLKNLIDWGNG SEQ ID NO: 1643 CGLFGEIEELIEEGLENLIDWANG SEQ ID NO: 1644 CGLFGEIEELIEEGLENLIDWSNG SEQ ID NO: 1645 CGLFGEIEELIEEGLENLIDW-(Aib)-NG SEQ ID NO: 1646 CGLFGEIEELIEEGLENLIDWPNG SEQ ID NO: 1647 CGLFGEIEELIEEGLENLIDWHNG SEQ ID NO: 1648 CGLFGEIEELIEEGLENLIDWQNG SEQ ID NO: 1649 CGLFGEIEELIEEGLENLIDWENG SEQ ID NO: 1650 CGLFEEIAELIEEGLENLIDWGNG SEQ ID NO: 1651 CELFEELAELLWEGLENLIDWGNS SEQ ID NO: 1652 CGLFGEIAELIWEGLENLIDWGNG SEQ ID NO: 1653 CGLLEEIEELLEEGLENLIDWGNS SEQ ID NO: 1654 CGLFAEIEELLEEGLENLIDWGNG SEQ ID NO: 1655 CGLL-(Aib)-EIEELLEEGLENLIDWWNS SEQ ID NO: 1656 CGLFGEIEELIEEGLENLIDWNNG SEQ ID NO: 1657 CGLFGEIEELIEEGLENLIDWDNG SEQ ID NO: 1658 CGLFGEIEELIEEGLENLIDWONG SEQ ID NO: 1659 CGLFAEIEELLEEGLENLIDWGNG SEQ ID NO: 1660 CGLL-Aib-EIEELLEEGLENLIDWGNS SEQ ID NO: 1661 CGLFGEIEELIEEGLENLIDGWNG SEQ ID NO: 1662 CGLFGEIEELIEEGLENLIDLWNG SEQ ID NO: 1663 CGWFGEIEELIEEGLENLIDWWNG SEQ ID NO: 1664 CGLFGEVEELIEEGLENLIDWWNG SEQ ID NO: 1665 CGLFGEIEEVIEEGLENLIDWWNG SEQ ID NO: 1666 CGLFGEIEELVEEGLENLIDWWNG SEQ ID NO: 1667 CGLFGEIEELAEEGLENLIDWWNG SEQ ID NO: 1668 CGLFGEIEELIDEGLENLIDWWNG SEQ ID NO: 1669 CGLFGEIEELIEDGLENLIDWWNG SEQ ID NO: 1670 CGLFGEIEELIEEGLEALIDWWNG SEQ ID NO: 1671 CGLFGEIEELIEEGLENIIDWWNG SEQ ID NO: 1672 CGLFGEIEELIEEGLEN-(Nle)-IDWWNG SEQ ID NO: 1673 CGLFGEIEELIEEGLENLIGWWNG SEQ ID NO: 1674 CGLFGEIEELIEEGLENLIDAWNG SEQ ID NO: 1675 CGLLEEIEELLEEGLENLIDWWNE SEQ ID NO: 1676 CELFEELAELLWEGLENLIDWWNE SEQ ID NO: 1677 CGLFGEIEELIEEGLENLIGWWNG SEQ ID NO: 1678 CGLFELIEGFIENGWEGMIDGWYGYGRKKRRQRR all (D) SEQ ID NO: 1679 CGLFEAIEGFIENGWEGMIDGWY all (D) SEQ ID NO: 1680 CGLFGEIEELIENGLKNLIDWWYGYGRKKRRQR all (D) SEQ ID NO: 1681 CGLFEALLELLESLWELLLEAYGRKKRRQRR all (D) SEQ ID NO: 1682 CGLFEEIEGFIENGWEGLIDWWYGYGHKKHHQHR all (D) SEQ ID NO: 1683 CGLFGEIEELIEEGLENLIDWWNE all (D) SEQ ID NO: 1684 CGLFGEIEELIEEGLENLIDWWNS all (D) SEQ ID NO: 1685 CGLFGEIEELIEEGLENLIDWWNQ all (D) SEQ ID NO: 1686 CYGRKKRRQRRLIRLWSHLIHIWFQNRRLKWKKK SEQ ID NO: 1687 CGLFEAIEEFIENGWEGMIDGWWYGYGRKKRRQRR SEQ ID NO: 1688 CGLFFAIEGFIENGWEGMIDWWYGYGRKKRRQRR ALL (D) SEQ ID NO: 1689 CGLFELIEGFIENGWEGMIDGWWYGYGRKKRRQRRK(STEARYL) ALL (D) SEQ ID NO: 1690 (STEARYL)GLFELIEGFIENGWEGMIDGWYGYGRKKRRQRRC ALL (D) SEQ ID NO: 1691 CFFGAIWEFIKSILK(STEARYL) ALL (D) SEQ ID NO: 1692 CGIFEAIAGLLKNIFKGIFEAIAGLLKNIFK ALL (D) SEQ ID NO: 1693 CIFGAIAGFIKNILKGLIDG ALL (D) SEQ ID NO: 1694 CGLFEAIEGFIENGWEGMIDGWWYGYGRKKRRQRRK(STEARYL) ALL(D) SEQ ID NO: 1695 (LAURYL)FFGAIWEFIKSILC ALL (D) SEQ ID NO: 1696

The D-amino acid, retro-inverso, and cysteine conjugation point variants of the peptides shown in Table 3 are also suitable.

The preferred peptides are listed in Table 4 below:

TABLE 4 Peptide Listing and ID Sequence SEQ ID CGLFEAIEGFIENGWEGMIDGWYGYGHKKHHQHH SEQ ID NO: 2 C-bAla-LFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 3 CGLFEAIEGFIEWGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 5 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQR SEQ ID NO: 7 CGLFHALLHLLHSLWHGLLHAWYGYGHKKHHQHR SEQ ID NO: 11 CGLFELIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 13 CGLFEAIEGFIENGWEG-Nle-IDGWYGYGRKKRRQRR SEQ ID NO: 19 CGLLEALEGLLESLWEGLLEAWYGYGRKKRRQRR SEQ ID NO: 22 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRRK(stearyl) SEQ ID NO: 27 CGLFEAIAGFIEGGWPGLINGWYGYGRKKRRQRRLHLLHHLLHHLHHL SEQ ID NO: 28 LHHLLHLLHHLLHHL CGLFEAIEGFIENGWEGMIDGWYGGGGLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 29 LHHLLHHL CGLFEAIEGFIENGWEGMIDGWYGLHLLHHLLHHLHHLLHHLLHL SEQ ID NO: 30 CGLFEALLELLESLWELLLEAYGRKKRRQRR SEQ ID NO: 31 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR SEQ ID NO: 32 CGLFHALLHLLHSLWHLLLHAWYGYGRKKRRQRR SEQ ID NO: 55 CGLFHALLHLLHSLWHLLLHAWYGYGHKKHHQHR SEQ ID NO: 56 CGIFGAIAGLLKNIFK SEQ ID NO: 63 CIFGAIAGFIKNIWKGLIDW SEQ ID NO: 64 stearyl-WEAALAEALAEALAEHLAEALAEALEALAAYGRKKRRQRRC SEQ ID NO: 69 CGFFHAFFHFFHSFWHGFFEA SEQ ID NO: 71 CGNFGEIEELIEEGLENLIDWWNG SEQ ID NO: 72 CFFGAIWEFIRNILEGF SEQ ID NO: 73 CFFGAIWEFIHSIL SEQ ID NO: 74 CGLFGEIEEFIENGWKGLIDWWYG SEQ ID NO: 86 CIFGIDDLIIGLLFVAIVEAGIGGYLLGSYGRKKRRQRR SEQ ID NO: 90 CFFGAIWEFIRSILK SEQ ID NO: 94 CFFGAIWEFIRSILE SEQ ID NO: 95 CGLFEAIEGFIENGWEGMIDWWYGYGRKKRRQRR SEQ ID NO: 106 CGLFEAIEGFIENGWEGMIDGWYGYGRKKRRQRR all (D) SEQ ID NO: 137 CRRQRRKKRGYGYWGDIMGEWGNEIFGEIAEFLG SEQ ID NO: 192 RRQRRKKRGYGYWGDIMGEWGNEIFGEIAEFLGC all(D) SEQ ID NO: 200 CRRQRRKKRGYGYWGDIMGEWGNEIFGEIAEFLG all(D) SEQ ID NO: 201 CGLFEAIEGFIENGWKGMIDGWYGYGRKKRRQRR SEQ ID NO: 228 CGLFEAIEGFIENGWKGMIDGWYGYGRKKRRQRR SEQ ID NO: 228 CGLFEAIEGFIENGWKGLIDWWYGYGRKKRRQRR SEQ ID NO: 266 CIFGAIAGFIKNIW SEQ ID NO: 283 CFFGAIWEFIRNIL SEQ ID NO: 333 FFGAIWEFIKSILC SEQ ID NO: 409 CFFGKIWEFIKSIL SEQ ID NO: 407 CFFGAIWEFAKSIL SEQ ID NO: 423 CGLFHALLHLLHSLWHLLLEA SEQ ID NO: 436 CGLFHALLHLLHSLWKLLLEW SEQ ID NO: 437 CGFFGEIAELIEEGLKGLIDWWNG SEQ ID NO: 461 CGLFGEIEELIEEGLENLIDWWNG SEQ ID NO: 462 CFFGAIWEFIHSIL all (D) SEQ ID NO: 463 CGIFEAIAGLLKSILKK(stearyl) SEQ ID NO: 468 CGIFGAIAGLLKSILKK(stearyl) SEQ ID NO: 469 CIFGAIAGFIKNILKGL all (D) SEQ ID NO: 470 CIFGAIAGFIKNILKGLK(stearyl) SEQ ID NO: 473 GLGKLINKIFGAIAGFIC all (D) SEQ ID NO: 474 CGLFGEIEELIEEGLENLIDWWNG all(D) SEQ ID NO: 491 CGNFGEIEELIEEGLENLIDWWNG all(D) SEQ ID NO: 492 CGFFGEIAELIEEGLKGLIDWWNG all(D) SEQ ID NO: 493 CGIFEAIAGLLKNIFK all(D) SEQ ID NO: 612 CIFGAIAGFIKNIWEGLI all (D) SEQ ID NO: 489 CGLFGEIEELIEEGLENLIDWGNG all (D) SEQ ID NO: 1074 CGLFGEIEELIEEGLENLIDWGNG SEQ ID NO: 1642 CGLFELIEGFIENGWEGMIDGWYGYGRKKRRQRR all (D) SEQ ID NO: 1679 CGLFEAIEGFIENGWEGMIDGWYG all (D) SEQ ID NO: 1680 CGLFGEIEELIENGLKNLIDWWYGYGRKKRRQRR all (D) SEQ ID NO: 1681 CGLFEALLELLESLWELLLEAYGRKKRRQRR all (D) SEQ ID NO: 1682 CGLFEEIEGFIENGWEGLIDWWYGYGHKKHHQHR all (D) SEQ ID NO: 1683 CGLFGEIEELIEEGLENLIDWWNE all (D) SEQ ID NO: 1684 CGLFGEIEELIEEGLENLIDWWNS all (D) SEQ ID NO: 1685 CGLFGEIEELIEEGLENLIDWWNQ all (D) SEQ ID NO: 1686 GFFGAIWEFIKSILC SEQ ID NO: 337

The D-amino acid, retro-inverso, and cysteine conjugation point variants of the peptides shown in Table 4 are also preferred.

Targeting Ligands

The modular compositions of the present invention may comprise a targeting ligand. In some embodiments, this targeting ligand may direct the modular composition to a particular cell. For example, the targeting ligand may specifically or non-specifically bind with a molecule on the surface of a target cell. The targeting moiety can be a molecule with a specific affinity for a target cell. Targeting moieties can include antibodies directed against a protein found on the surface of a target cell, or the ligand or a receptor-binding portion of a ligand for a molecule found on the surface of a target cell. Examples and a further discription of targeting ligands can be found in WO2009/126933, which is hereby incorporated by reference.

The targeting ligands are selected from the group consisting of an antibody, a ligand-binding portion of a receptor, a ligand for a receptor, an aptamer, D-galactose, N-acetyl-D-galactose (GalNAc), multivalent N-acytyl-D-galactose, D-mannose, cholesterol, a fatty acid, a lipoprotein, folate, thyrotropin, melanotropin, surfactant protein A, mucin, carbohydrate, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, N-acetyl-glucosamine, multivalent mannose, multivalent fructose, glycosylated polyaminoacids, transferin, bisphosphonate, polyglutamate, polyaspartate, a lipophilic moiety that enhances plasma protein binding, a steroid, bile acid, vitamin B12, biotin, an RGD peptide, an RGD peptide mimic, ibuprofen, naproxen, aspirin, folate, and analogs and derivatives thereof.

The preferred targeting ligands are selected from the group consisting of D-galactose, N-acetyl-D-galactose (GalNAc), GalNAc2, and GalNAc3, cholesterol, folate, and analogs and derivatives thereof.

Lipids

Lipophilic moieties, such as cholesterol or fatty acids, when attached to highly hydrophilic molecules such as nucleic acids can substantially enhance plasma protein binding and consequently circulation half life. In addition, lipophilic groups can increase cellular uptake. For example, lipids can bind to certain plasma proteins, such as lipoproteins, which have consequently been shown to increase uptake in specific tissues expressing the corresponding lipoprotein receptors (e.g., LDL-receptor or the scavenger receptor SR-B1). Lipophilic conjugates can also be considered as a targeted delivery approach and their intracellular trafficing could potentially be further improved by the combination with endosomolytic agents.

Exemplary lipophilic moieties that enhance plasma protein binding include, but are not limited to, sterols, cholesterol, fatty acids, cholic acid, lithocholic acid, dialkylglycerides, diacylglyceride, phospholipids, sphingolipids, adamantane acetic acid, 1-pyrene butyric acid, dihydrotestosterone, 1,3-Bis-O(hexadecyl)glycerol, geranyloxyhexyl group, hexadecylglycerol, borneol, menthol, 1,3-propanediol, heptadecyl group, palmitic acid, myristic acid, O3-(oleoyl)lithocholic acid, O3-(oleoyl)cholenic acid, dimethoxytrityl, phenoxazine, aspirin, naproxen, ibuprofen, vitamin E and biotin etc. Examples and a further discription of lipids can be found in WO2009/126933, which is hereby incorporated by reference.

The preferred lipid is cholesterol.

Solubilizing Agents

The modular composition may comprise one or more other moieties/ligands that may enhance aqueous solubility, circulation half life and/or cellular uptake. These can include naturally occurring substances, such as a protein (e.g., human serum albumin (HSA), low-density lipoprotein (LDL), high-density lipoprotein (HDL), or globulin); or a carbohydrate (e.g., a dextran, pullulan, chitin, chitosan, inulin, cyclodextrin or hyaluronic acid). These moieties may also be a recombinant or synthetic molecule, such as a synthetic polymer or synthetic polyamino acids. Examples include polylysine (PLL), poly L-aspartic acid, poly L-glutamic acid, styrene-maleic acid anhydride copolymer, poly(L-lactide-co-glycolied) copolymer, divinyl ether-maleic anhydride copolymer, N-(2-hydroxypropyl)methacrylamide copolymer (HMPA), polyethylene glycol (PEG, e.g., PEG-0.5K, PEG-2K, PEG-5K, PEG-10K, PEG-12K, PEG-15K, PEG-20K, PEG-40K), methyl-PEG (mPEG), [mPEG]2, polyvinyl alcohol (PVA), polyurethane, poly(2 ethylacryllic acid), N-isopropylacrylamide polymers, or polyphosphazine. Examples and a further discription of solubilizing agents can be found in WO2009/126933, which is hereby incorporated by reference.

The preferred solubilizing group is PEG 0.5K to 30K.

Method of Treatment

In one aspect, the invention features, a method of treating a subject at risk for or afflicted with a disease that may benefit from the administration of the modular composition of the invention. The method comprises administering the modular composition of the invention to a subject in need thereof, thereby treating the subject. The oligonucleotide that is administered will depend on the disease being treated. See WO2009/126933 for additional details regarding methods of treatments for specific indications.

Formulation

There are numerous methods for preparing conjugates of oligonucleotide compounds. The techniques should be familiar to those skilled in the art. A useful reference for such reactions is Bioconjugate Techniques, Hermanson, G. T., Academic Press, San Diego, Calif., 1996. Other references include WO2005/041859; WO2008/036825 and WO2009/126933.

EXAMPLES

The invention is further illustrated by the following examples, which should not be construed as further limiting. The contents of all references, pending patent applications and published patents, cited throughout this application are hereby expressly incorporated by reference. The siRNAs described herein were designed to target the ubiquitously expressesd gene SSB (Sjogren syndrome antigen B; NM_009278.4).

Linker groups may be connected to the oligonucleotide or siRNA strand(s) at a linkage attachment point (LAP) and may include any carbon-containing moiety, in some embodiments having at least one oxygen atom, at least one phosphorous atom, and/or at least one nitrogen atom. In some embodiments, the phosphorous atom forms part of a terminal phosphate, or phosphorothioate, group on the linker group, which may serve as a connection point for the oligonucleotide strand. In certain embodiments, the nitrogen atom forms part of a terminal ether, ester, amino or amido (NHC(O)—) group on the linker group, which may serve as a connection point for the linkers of interest, endosomolytic unit, cell penetrating peptide, solubilizing group, lipid, targeting group, or additional linkers of interest. These terminal linker groups include, but are not limited to, a C₆ hexyl, C₅ secondary-hydroxy, C₃ thiol or C₆ thiol moiety. An example from the RNA sequences described below is C6 hexyl: [(CH₂)₆ NH₂].

The siRNA sequences described in the Examples herein are shown in Table 5.

TABLE 5 Sequence SEQ ID Entry Code Compound stand Sequence NO:  1 b CTNNB1 passenger [6amiL][iB][omeC][omeU][clickG][omeU][omeU][fluG] 1697 [fluG][fluA][omeU][omeU][fluG][fluA][omeU][omeU] [omeC][fluG][clickA][fluA][fluA][omeU][omeU][iB] [C3SH] CTNNB1 guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1698 [omeC][fluA][omeA][fluU][omeC][fluC][omeA][fluA] [omeC][fluA][omeG][omeUs][omeU]  2 c ApoB passenger [C6SH][iB][omeC][omeU][omeU][omeU][fluA][fluA][omeC] 1699 [fluA][fluA][omeU][omeU][omeC][omeC][omeU][fluG] [fluA][fluA][fluA][omeU][dTs][dT][iB][6amiL] ApoB guide [rAs][rUs][rUs][omeU][omeC][fluA][fluG][fluG][fluA] 1700 [fluA][omeU][omeU][fluG][fluU][omeU][fluA][fluA] [fluA][fluG][omeUs][omeU]  3 d CTNNB1 passenger [6amiL][iB][omeC][omeU][clickG][omeU][omeU][fluG] 1701 [gluG][fluA][omeU][omeU][fluG][fluA][omeU][omeU] [omeC][fluG][clickA][fluA][fluA][omeUs][omeU] [iB][C3SH] guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1702 [omeC][fluA][omeA][fluU][omeC][fluC][omeA][fluA] [omeC][fluA][omeG][omeUs][omeU]  4 e CTNNB1 passenger [6amiL][iB][omeC][omeU][fluG][omeU][omeU][fluG][gluG] 1703 [fluA][omeU][omeU][fluG][fluA][omeU][omeU][omeC] [fluG][clickA][fluA][fluA][omeUs][omeU][iB][C3SH] guide [omeUs][fluAs][omeUs][fluC][omeG][fluA][omeA][fluU] 1704 [omeC][fluA][omeA][fluU][omeC][fluC][omeA][fluA] [omeC][fluA][omeG][omeUs][omeU]  5 f CTNNB1 passenger [6amiL][iB][omeC][omeU][clickG][omeU][omeU][fluG] 1705 [gluG][fluA][omeU][omeU][fluG][fluA][clickU][omeU] [omeC][fluG][fluA][clickA][fluA][omeUs][omeU][iBC] [C3SHSup] guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1706 [omeC][fluA][omeA][fluU][omeC][fluC][omeA][fluA][omeC] [fluA][omeG][omeUs][omeU]  6 g CTNNB1 passenger [6amiL][iB][omeC][omeU][clickG][omeU][omeU][fluG] 1701 [fluG][clickA][omeU][omeU][fluG][fluA][clickU][omeU] [omeC][fluG][fluA][clickA][fluA][omeUs][omeU][iB] [C3SHSup] guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1708 [omeC][fluA][omeA][fluU][omeC][fluC][omeA][fluA] [omeC][fluA][omeG][omeUs][omeU]  7 h CTNNB1 passenger [LiCholinker][iB][omeC][omeU][fluG][omeU][omeU][fluG] 1709 [fluG][fluA][omeU][omeU][fluG][fluA][omeU][omeU][omeC] [fluG][fluA][fluA][fluA][omeUs][omeU][iB][6amiL] guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1710 [omeC][fluA][omeA][fluU][omeC][fluC][omeA][fluA] [omeC][fluA][omeG][omeUs][omeU]  8 i CTNNB1 passenger [amino modifier C2 1711 dT]][iB][omeC][omeU][clickG][omeU][omeU][fluG][fluG] [gluA][omeU][omeU][fluG][fluA][omeU][omeU][omeC] [fluG][clickA][fluA][fluA][omeUs][omeU][iB][C3SSC3OH] guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1712 [omeC][fluA][omeA][fluU][omeC][fluC][omeA][fluA] [omeC][fluA][omeG][omeUs][omeU]  9 j CTNNB1 passenger [6amiL][iB][omeC][omeU][clickG][omeU][omeU][fluG] 1713 [gluG][fluA][omeU][omeU][fluG][fluA][omeU][omeU][omeC] [fluG][fluA][fluA][fluA][omeUs][omeU][iB][C3SH] guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1714 [omeC][clickA][omeA][fluU][omeC][fluC][clickA][fluA] [omeC][fluA][omeG][omeUs][omeUSup] 10 k CTNNB1 passenger [6amiL][iB][omeC][omeU][clickG][omeU][omeU][fluG] 1715 [gluG][fluA][omeU][omeU][fluG][fluA][omeU][omeU][omeC] [fluG][fluA][fluA][fluA][omeUs][omeU][iB][C3SH] guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1716 [omeC]][fluA][omeA][fluU][omeC][fluC][clickA][fluA] [omeC][fluA][omeG][omeUs][omeU] 11 l CTNNB1 passenger [6amiL][iB][omeC][omeU][fluG][omeU][omeU][fluG][fluG] 1717 [gluA][omeU][omeU][fluG][fluA][omeU][omeU][omeC][fluG] [fluA][fluA][fluA][omeUs][omeU][iB][6amiL] guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1718 [omeC]][fluA][omeA][fluU][omeC][fluC][clickA][fluA] [omeC][fluA][omeG][omeUs][omeU] 12 m CTNNB1 passenger [6amiL][iB][omeC][omeU][clickG][omeU][omeU][fluG] 1719 [fluG][fluA][omeU][omeU][fluG][fluA][clickU][omeU] [omeC][fluG][fluA][clickA][fluA][omeUs]omeU][iB] [C3SHSup] guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1720 [omeC]][fluA][omeA][fluU][omeC][fluC][clickA][fluA] [omeC][fluA][omeG][omeUs][omeU] As used herein, ome = 2′ methoxy; flu = 2′ fluoro; click = 2′ propagyl; iB = inverted abasic; ″s″ subscript = phosphorothioate; and r = 2′ ribo; 6 amil = n-hexylamino; C3SH = n-propylthiol; and C6SH = n-hexythiol.

Preparations of tetraGalNAc ligands and tetraGalNAc-siRNA conjugates are described below in the examples and synthetic schemes. Note that the siRNA depictions below are for illustrative purposes. Specific sequence information can be found in Table 5.

Section A Examples 1-2 Synthesis of TetraGalNAc Ligand Compounds A9 and A10

The following Scheme 1 was used to prepare TetraGalNAc Compounds 9 and 10.

Synthesis of (2S)-2, 6-bis[bis (prop-2-yn-1-yl)amino]hexanoic acid (Compound A1)

Into a 2000-mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of (2S)-2,6-diaminohexanoic acid (50 g, 342.03 mmol, 1.00 equiv) in acetonitrile (1000 mL) and heated to 50° C. To this was added potassium hydroxide (22.6 g, 0.4025 mol, 1.00 equiv, 85%). The resulting solution was stirred for 30 min. Then 3-bromoprop-1-yne (29.5 mL, 1.00 equiv) was added. The resulting solution was stirred for 1 hour at 50° C. additional potassium hydroxide (22.6 g, 0.4025 mol, 1.00 equiv) was added to the solution and stirred for 30 min at 50° C. To this was added 3-bromoprop-1-yne (29.5 mL, 1.00 equiv). The resulting solution was stirred for 1 hour. To this was added potassium hydroxide (22.6 g, 0.4025 mol, 1.00 equiv) again. The resulting solution was stirred for 30 min at 50° C., followed by addition of more 3-bromoprop-1-yne (29.5 mL, 1.00 equiv). The resulting solution was stirred for 1 hour. To this was added potassium hydroxide (22.6 g, 0.4025 mol, 1.00 equiv). The resulting solution was stirred for 30 min. To this was added 3-bromoprop-1-yne (29.5 mL, 1.00 equiv). The resulting solution was stirred for 3 hours. The reaction mixture was cooled to 25° C. with a water/ice bath. The solid was filtered out. The filtrate was adjusted to pH 4 with HCl (6M). The solid was filtered out. The filtrate was concentrated under vacuum.

The residue was applied onto a silica gel column and eluted with dichloromethane/methanol (100:1-25:1). This resulted in (2S)-2, 6-bis[bis (prop-2-yn-1-yl)amino]hexanoic acid (Compound A1) as a light yellow oil.

MS (ES, m/z): 297.2, [M−H]-¹HNMR(CDCl₃, 500 MHz, ppm): 3.62 (d, J=2.0 Hz, 4H), 3.52-3.49 (m, 1H), 3.50 (d, J=2.4 Hz, 4H), 2.62 (t, J=7.1 Hz, 2H), 2.30 (t, J=2.4 Hz, 2H), 2.27 (t, J=2.4 Hz, 2H), 1.88-1.79 (m, 2H), 1.60-1.53 (m, 2H), 1.52-1.43 (m, 2H).

Synthesis of 2-(2-hydroxyethoxy)ethyl 4-methylbenzenesulfonate (Compound A3)

Into a 2000-mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of 2-(2-hydroxyethoxy)ethan-1-ol (A2, 42.4 g, 399.55 mmol, 1.00 equiv) in dichloromethane (1000 mL) and triethylamine (27.9 g, 275.72 mmol, 0.25 equiv). To the above was added p-toluenesulfonyl chloride (19.1 g, 100.18 mmol, 0.50 equiv). After stirred for 1 h at 25° C., the resulting mixture was washed with 1×500 mL of aq. potassium hydrosulfate (1M) and 1×500 mL of aq. sodium bicarbonate (5%) respectively. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column and eluted with dichloromethane/methanol (100:1). This resulted in 2-(2-hydroxyethoxy)ethyl 4-methylbenzenesulfonate (Compound A3) as a colorless oil.

Synthesis of 2-(2-azidoethoxy)ethan-1-ol (Compound A4)

Into a 500-mL round-bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of 2-(2-[[(4-2-(2-hydroxyethoxy)ethyl 4-methylbenzenesulfonate (A3, 50 g, 192.08 mmol, 1.00 equiv) in N,N-dimethylformamide (250 mL). This was followed by the addition of sodium azide (18.79 g, 289.03 mmol, 1.50 equiv) at 25° C. The resulting solution was stirred for 5 h at 100° C. in an oil bath. The reaction mixture was cooled and filtered. The filtrate was concentrated under vacuum. The residual solution was diluted with 1000 mL of dichloromethane and washed with 1×500 mL of water. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column and eluted with dichloromethane/methanol (80:1). This resulted in 2-(2-azidoethoxy)ethan-1-ol (Compound A4) as a colorless oil.

¹HNMR (CDCl₃, 400 MHz, ppm): 3.42-3.45 (t, J=4.8 Hz, 2H), 3.63-3.65 (t, J=4.8 Hz, 2H), 3.71-3.74 (t, J=4.8 Hz, 2H), 3.71-3.79 (m, 2H).

Synthesis of (3R,4R,5R,6R)-3-acetamido-6-(acetoxymethyl)tetrahydro-2H-pyran-2,4,5-triyl triacetate (Compound A6)

Into a 2000-mL 3-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of (3R,4R,5R,6R)-3-amino-6-(hydroxymethyl)tetrahydro-2H-pyran-2,4,5-triol hydrochloride (A5, 120 g, 556.50 mmol, 1.00 equiv) in pyridine (1200 mL). This was followed by the addition of acetic anhydride (341.6 g, 3.35 mol, 6.00 equiv) dropwise with stirring at 0° C. The resulting solution was stirred overnight at 25° C. The reaction was then quenched by the addition of 8000 mL of water/ice. The solid was collected by filtration. This resulted in (3R,4R,5R,6R)-3-acetamido-6-(acetoxymethyl)tetrahydro-2H-pyran-2,4,5-triyl triacetate (Compound A6) as a white solid.

Synthesis of (3aR,5R,6R,7R,7aR)-5-(acetoxymethyl)-2-methyl-5,6,7,7a-tetrahydro-3aH-pyrano[3,2-d]oxazole-6,7-diyl diacetate (Compound A7)

Into a 2000-mL round-bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of (3R,4R,5R,6R)-3-acetamido-6-(acetoxymethyl)tetrahydro-2H-pyran-2,4,5-triyl triacetate (A6, 30 g, 77.05 mmol, 1.00 equiv) in dichloromethane (1500 mL), then added iron (III) chloride (30 g, 184.95 mmol, 2.40 equiv). The resulting mixture was stirred for 2 h at 25° C. The reaction was then quenched by the addition of 1000 mL of water/ice. The organic layer was washed with 1×1000 mL of sodium aq. bicarbonate and 1×1000 mL of water, dried over anhydrous sodium sulfate and concentrated under vacuum. This resulted in (3aR,5R,6R,7R,7aR)-5-(acetoxymethyl)-2-methyl-5,6,7,7a-tetrahydro-3 aH-pyrano[3,2-d]oxazole-6,7-diyl diacetate (Compound A7) as yellow oil.

¹HNMR(CDCl₃, 300 MHz, ppm): 2.03 (s, 9H), 2.12 (s, 3H), 3.97-4.27 (m, 4H), 4.90-4.93 (m, J=3.3 Hz, 1H), 5.45-5.47 (t, J=3.0 Hz, 1H), 5.98-6.00 (d, J=6.6 Hz, 1H).

Synthesis of (2R,3R,4R,5R,6R)-5-acetamido-2-(acetoxymethyl)-6-[2-(2-azidoethoxy)ethoxy]tetrahydro-2H-pyran-3,4-diyl diacetate (Compound A8)

Into a 500-mL round-bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of (3aR,5R,6R,7R,7aR)-5-(acetoxymethyl)-2-methyl-5,6,7,7a-tetrahydro-3aH-pyrano[3,2-d]oxazole-6,7-diyl diacetate (A7, 40 g, 121.47 mmol, 1.00 equiv) in 1,2-dichloroethane (200 mL), 2-(2-azidoethoxy)ethan-1-ol (A4, 23.89 g, 182.18 mmol, 1.50 equiv). To the above several 4A zeolite was added. The resulting mixture was stirred for 1 h at 25° C. Then trimethylsilyl trifluoromethanesulfonate (10.8 mL, 0.50 equiv) was added. After stirred overnight at 25° C., the reaction mixture was diluted with 500 mL of dichloromethane and washed with 1×500 mL of water, 1×500 mL of aq. sodium bicarbonate and 1×500 mL of water. The organic layer was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column and eluted with dichloromethane/methanol (100:1). This resulted in (2R,3R,4R,5R,6R)-5-acetamido-2-(acetoxymethyl)-6-[2-(2-azidoethoxy)ethoxy]tetrahydro-2H-pyran-3,4-diyl diacetate (A8) as a colorless oil.

MS (m/z): 461.1, [M+H]⁺

¹HNMR(CDCl₃, 500 MHz, ppm) 5.78 (d, J=8.90 Hz, 1H), 5.36 (d, J=2.9 Hz, 1H), 5.22 (dd, J=11.2, 3.6 Hz, 1H), 4.77 (d, J=8.3 Hz, 1H), 4.19-4.12 (m, 2H), 4.11-4.05 (m, 1H), 3.98-3.92 (m, 2H), 3.82-3.78 (m, 1H), 3.71-3.63 (m, 4H), 3.49-3.38 (m, 2H), 2.16 (s, 3H), 2.05 (s, 3H), 2.01 (s, 3H), 1.97 (s, 3H).

Synthesis of (S)-2,6-bis(bis((1-(2-(2-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-diacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)-1H-1,2,3-triazol-4-yl)methyl)amino)hexanoic acid (Compound A9, tetraGalNAc Acetate) (A9) (Ex. 1)

Into a 250-mL round bottom flask purged and maintained with an inert atmosphere of nitrogen was charged (2S)-2, 6-bis [bis (prop-2-yn-1-yl) amino]hexanoic acid (A1, 1.0 g, 1.0 equiv), (2R,3R,4R,5R,6R)-5-acetamido-2-(acetoxymethyl)-6-[2-(2-azidoethoxy)ethoxy]tetrahydro-2H-pyran-3,4-diyl diacetate (A8, 9.26 g, 6.0 equiv), anhydrous THF 50 mL, CuBrSMe₂ (0.138 g, 0.20 equiv), and anhydrous DBU (1.5 ml, 3.0 equiv) in respective order. The resulting solution was stirred for 16 h at room temperature, quenched with acetic acid (0.75 mL, 4.0 equiv), treated with MP-TMT resin (Part No: 801472, from Biotage) (9 g), aged at room temperature for 16 h, filtered, and concentrated the filtrate to a foam solid. The solid was then dissolved in CH₂Cl₂ (140 mL), and washed with AcOH/NaCl solution (140 mL). The AcOH/NaCl solution was prepared with 1 mL AcOH and 100 mL 20% NaCl solution. The bottom organic layer was concentrated, and purified on a SiO₂ column (220 g), eluting with CH₂Cl₂/MeOH. This resulted in (S)-2,6-bis(bis((1-(2-(2-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-diacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)-1H-1,2,3-triazol-4-yl)methyl)amino)hexanoic acid (Compound A9) as a white solid.

MS (m/z): 2139.5, [M+H]⁺

Synthesis of (S)-2,6-bis(bis((1-(2-(2-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)-1H-1,2,3-triazol-4-yl)methyl)amino)hexanoic acid (Compound A10, TetraGalNAc) (A10) (Ex. 2)

Into a 250-mL round-bottom flask purged and maintained with an inert atmosphere of nitrogen was charged (S)-2,6-bis(bis((1-(2-(2-(((2R,3R,4R,5R,6R)-3-acetamido-4, 5-diacetoxy-6-(acetoxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)-1H-1,2,3-triazol-4-yl)methyl)amino)hexanoic acid (A9, 6.9 g, 1.0 equiv), Na₂CO₃ (6.83 g, 20 eq), water (56 mL), and MeOH (32 mL) in respective order. The reaction was aged at room temperature for 16 h, concentrated to residue, redissolved in water (50 mL), and purified on Combiflash C18 gold reverse column (415 g), eluting with water/MeCN. After concentration under vacuum, the product was dissolved in minimum amount of water, and lyophilized to obtain (S)-2,6-bis(bis((1-(2-(2-(((2R,3R,4R,5R,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)-1H-1,2,3-triazol-4-yl)methyl)amino)hexanoic acid (Compound A10) as a white solid.

MS (m/z): 1657 [M+Na]⁺

¹HNMR (D₂O, 500 MHz, ppm): 8.05 (s, 2H), 7.91 (s, 2H), 4.62 (t, J=5.0 Hz, 4H), 4.57 (t, J=5.0 Hz, 4H), 4.45-4.41 (d, J=8.6 Hz, 4H), 3.99-3.82 (m, 28H), 3.80-3.61 (m, 28H), 3.14 (t, J=7.1 Hz, 1H), 2.52 (broad s, 2H), 1.99 (s, 6H), 1.98 (s, 6H), 1.73 (m, 2H), 1.60 (m, 2H), 1.29 (m, 2H).

Section B Preparation of B2 to B5 Examples 3-6

Scheme 2 as shown in FIG. 5A-1 to FIG. 5D, was used to prepare B Conjugates (Ex. 3-6).

Synthesis of B2 (Ex. 3)

A10 (86 mg, 0.053 mmol) and DIEA (57.6 μL, 0.330 mmol) were dissolved in DMSO (500 μL), then added to a solution of HATU (301 μL, 0.079 mmol) and stirred for 15 min. Starting material passenger strand B1 (101 mg, 0.013 mmol) was dissolved in water (168 μL) and DMSO (1.5 mL). The HATU solution was added to the RNA solution and aged for 15 min. The reaction mixture was diluted with water (50 mL) and centrifugal dialyzed three times against water over a 3 k membrane. The concentrate was loaded onto an HPLC fitted with a Dionix ProPac SAX 22×250 mm column. The product was gradient eluted starting at 95% A (2:3 H₂O:2,2,2-trifluoroethanol, 20 mM TEA) up to 40% solvent B (2:3 H₂O:2,2,2-trifluoroethanol, 20 mM TEA, 1M CsCl). The fractions were diluted with water to reduce the 2,2,2-trifluoroethanol content to 25% and centrifugal dialyzed three times against water over a 3 k membrane. The concentrate was freeze dried to afford the product as a white amorphous solid.

Expected mass: 9267.5, found mass: 9267.0

Synthesis of B3 (Ex. 4)

To a solution of B2 (606 mg, 0.065 mmol) in water (32 mL) was added TEAA (1.64 mL, 2M), aqueous DTT (0.65 mL, 1M), and TEA (0.65 mL, 4.69 mmol). The reaction mixture was aged for 10 min. The reaction mixture was then diluted with water and centrifugal dialyzed three times against water over a 3 k membrane. The concentrate was taken forward without further isolation. Expected mass: 9177.4, found mass: 9179.0

Synthesis of B4 (Ex. 5)

To a solution of B3 (350 mg, 0.038 mmol) in water (3 mL) was added N-(2-aminoethyl)-maleimide trifluoroacetate salt (194 mg, 0.763 mmol). The reaction mixture was aged for 30 min, after which it was purified by RP-HPLC (95:5-5:95% A:B linear gradient (A=100 mM aqueous TEAA; B=100 mM TEAA in acetonitrile) Waters Phenyl xbridge Column. Fractions containing B4 were centrifugal dialyzed three times against water over a 3 k membrane and the concentrate was lyophilized to give product as a white amorphous solid.

Synthesis of B5 (Ex. 6)

To a solution of B4 (286 mg, 0.031 mmol) in aqueous sodium bicarbonate (3.0 mL, 200 mM) was added a solution of NHS-dPEG12-SPDP (280 mg, 0.307 mmol) in acetonitrile (0.5 mL). The reaction mixture was aged for 30 min, after which it was treated with aqueous TEAA (1.0 mL, 2M) and purified by RP-HPLC (95:5-5:95% A:B linear gradient (A=100 mM aqueous TEAA; B=100 mM TEAA in acetonitrile) Waters Phenyl xbridge Column. Fractions containing B5 were centrifugal dialyzed three times against water over a 3K membrane and the concentrate was lyophilized to give product as a white amorphous solid. Measured mass=10117

Examples 7-8 Preparation of B6-Seq32

Scheme 3 as shown in FIG. 6A to FIG. 6B was used to prepare Conjugates B6-P32 and B8-seq32 (Ex. 7-8).

Synthesis of Conjugate B6-seq32 (Ex. 7)

B5 (50 mg, 5 umol, 1 eq.) was dissolved in 50 mM AcOH in 2,2,2-trifluoroethanol (5 mL). Peptide Seq32 (51 mg, 13 umol, 2.5 eq.) was dissolved in guanidine-HCl (8M, 500 uL), diluted with 50 mM AcOH in 2,2,2-trifluoroethanol (5 mL). The peptide solution was added dropwise to the stirring RNA solution over 5 min, and the reaction was left at room temperature for 1 hour. The reaction was diluted with formamide (10 mL), and 1.5 mL aliquots of the reaction mixture were loaded onto an HPLC fitted with a Dionex ProPac SAX-10 22×250 mm column. The product was gradient eluted starting at 98% solvent A (2:3 H₂O:2,2,2-trifluoroethanol, 40 mM TEA) up to 35% solvent B (2:3 H₂O:2,2,2-trifluoroethanol, 40 mM TEA, 1M guanidine-HCl) over 10 min at 20 mL/min. The fractions were diluted with water to reduce the 2,2,2-trifluoroethanol content to 25% and centrifugal dialyzed three times against water over a 10 k membrane. The concentrate was freeze dried to afford the product as a white amorphous solid. Expected mass: 13961.9, found mass: 13962.0

Synthesis of Conjugate B8-seq32-b (Ex. 8)

Guide strand (B7, 17.7 mg) was dissolved in water (5 mL) and added to a vial containing B6-seq 42 (36.2 mg). The solution was thoroughly mixed and left at room temperature for 2 hours. The solution was freeze dried to afford the duplex as a white amorphous solid.

Synthesis of Additional B8-Peptide Conjugates

Additional conjugates of B8 and Peptide Sequence and duplexes were prepared in a manner analogous to that used for B8-seq32-b.

Examples 9-11 Preparation of B9 and B10-seq32 and 11-seq32

Scheme 4 as shown in FIG. 7A, FIG. 7B and FIG. 7C was used to prepare B9, B10-seq32 and B11-seq32.

Synthesis of B9 (Ex. 9)

Compound B3 (120 mg, 0.0132 mmol) in water (5 mL) was added dropwise to a stirring solution of 2,2′-dipyridyldisulfide (29 mg, 0.132 mmol, 10 eq.) dissolved in methanol (5 mL). The solution was diluted with water to bring the methanol content to 20% and centrifugal dialyzed three times against water over a 3K membrane. The concentrate was freeze dried to afford the product as an amorphous white solid. Expected mass: 9166.5, found mass: 9165.5

Synthesis of B10-seq32 (Ex. 10)

B9 (15 mg, 1.615 umol) was dissolved in water (150 uL) and was diluted with 50 mM AcOH in TFE (1.5 mL). In a separate vial, P32 (8.79 mg, 2.155 umol) was dissolved in 8 M guanidine HCl (60 uL) and diluted with 50 mM AcOH in TFE (1.5 mL), then added to the RNA solution. The reaction mixture was aged for 15 min, then was diluted with formamide and purified by AEX (95:5-55:45 A:B linear gradient (A=20 mM TEA in 60% aqueous TFE; B=1M CsCl and 20 mM TEA in 60% aqueous TFE), Dionix Propac column. Fractions containing B10-Seq 32 were centrifugal dialyzed three times against water over a 10K membrane and the concentrate was lyophilized to give product as a white amorphous solid.

Synthesis of B11-seq32-b (Ex. 11)

B10-seq 32 (9.68 mg, 0.730 umol) was treated with a solution of B7 (5.00 mg, 0.730 umol) dissolved in PBS (500 uL) and aged for 30 min. Excess guide strand was removed by AEX purification (95:5-55:45 A:B linear gradient (A=20 mM TEA in 60% aqueous TFE; B=1M CsCl and 20 mM TEA in 60% aqueous TFE), Dionix Propac column. Fractions containing B11-seq 32 were centrifugal dialyzed three times against water over a 10K membrane and the concentrate was lyophilized to give product as a white amorphous solid.

Examples 12-14 Additional Synthesis of B11-Peptide Conjugates

Additional conjugates of B11 and peptide sequences and corresponding duplexes were prepared in a manner analogous to that used for B11-seq32-b.

Scheme 5 is shown in FIG. 7D, FIG. 7E and FIG. 7F.

Synthesis of B12 (Ex. 12)

B3 (50 mg, 5.4 μmol) was dissolved in water (3 mL, ˜17 mg/mL) and Compound 1, 1,1′-(ethane-1,2-diyl)bis(1H-pyrrole-2,5-dione), (16 mg, 0.073 mmol) was dissolved in DMF (1.2 mL) in separate vials. The B3 solution was added to Compound 1 solution and stirred for 10 min. The reaction was diluted with water to 15 mL and then dialyzed 4 times on 3 K MWCO membrane against water. The reaction was then filtered (0.22 m syringe filter) and lyophilzed to afford a white solid, B12. Expected mass: 9397.535. Observed mass: 9400.0.

Synthesis of B12-seq13 (Ex. 13)

See Synthesis of B10-seq32 for reaction procedure.

B12-seq13. Expected mass: 13518.215

Synthesis of B13-seq13-b (Ex. 14)

See Synthesis of B11-seq32 for reaction procedure.

B13-seq13-b. Expected mass: 20370.215

Additional Synthesis of B13-Peptide Conjugates

Additional conjugates of B13 and peptide sequences were prepared in a manner analogous to that used for B13-seq13.

Examples 15-16 Preparation of B15-seq32 and B16-seq32-b

Scheme 6 as shown in FIG. 7G-1 to FIG. 7G-2 was used to prepare B16-seq32 and B17-seq32-b.

Synthesis of B14

B3 (100 mg, 10.9 μmol) was dissolved in water (10 mL) and dioxane (20 mL) was treated with bis maleimide dissolved in dioxane (3.8 mL) to give a cloudy mixture. The reaction was stirred for 1.5 hours, after which it was quenched with N-methylmaleimide (36.3 mg, 0.327 mmol). The reaction mixture was diluted with water and centrifugal dialyzed once against water over a 3 k membrane. The concentrate was filtered and purified by RP-HPLC (95:5-5:95% A:B linear gradient (A=100 mM aqueous TEAA; B=100 mM TEAA in acetonitrile) Waters Phenyl xbridge Column). Fractions containing product were dialyzed and lyophilized to give B14 as an amorphous white powder. Measured mass=9531

Synthesis of B15-seq 32 (Ex. 15)

B14 (5 mg, 0.524 μmol) was dissolved in formamide solution (2M thiourea, 50 mM MES buffer at pH 6.5, 500 μL). In a separate vial, peptide sequence 32 (4.28 mg, 1.048 mol) was dissolved in formamide solution (2M thiourea, 50 mM MES buffer at pH 6.5, 500 μL), then was added to the RNA solution. After aging one hour at room temperature, the reaction mixture was loaded onto an HPLC fitted with a Dionex ProPac SAX-10 22×250 mm column. The product was gradient eluted starting at 98% solvent A (2:3 H₂O:2,2,2-trifluoroethanol, 40 mM TEA) up to 35% solvent B (2:3 H₂O:2,2,2-trifluoroethanol, 40 mM TEA, 1M guanidine-HCl) over 10 min at 20 mL/min. The fractions were diluted with water to reduce the 2,2,2-trifluoroethanol content to 25% and centrifugal dialyzed three times against water over a 10 k membrane. The concentrate was freeze dried to afford the product as a white amorphous solid.

Synthesis of B16-seq32-b (Ex. 16)

B15-seq 32 (2.11 mg, 0.155 μmol) was treated with a solution of B7 (1.062 mg, 0.155 μmol) in water (212 μL) and aged at room temperature for 2 hours. The solution was lyophilized to give the product as a white amorphous solid.

Section C Examples 17-21 Preparation of C1 to C3, C4-seq32 and C6-seq32

Scheme 7 as shown in FIG. 8A to FIG. 8D was used to prepare C1 to C3, C4-seq32 and C6-seq32.

Synthesis of C1 (Ex. 17)

1,2-Diaminododecane (100 mg, 0.499 mmol) was dissolved in chloroform (3.3 mL) and cooled to 0° C., then treated with N-methoxycarbonyl-maleimide (234 mg, 1.50 mmol) and tetrabutylammonium hydrogen sulfate (170 mg, 0.499 mmol). DIPEA (209 uL, 1.20 mmol) was slowly added and the reaction aged for 10 minutes at 0° C. The ice bath was removed and the reaction was treated with aqueous saturated sodium bicarbonate solution (6.6 mL). After aging 3.5 hours at room temperature, the reaction mixture was extracted with ethyl acetate (3×15 mL). The combined organic layers were dried with sodium sulfate and then solvent removed in vacuo. The crude product was purified by flash chromatography with a 100:0-0:100% A:B linear gradient (A=hexanes; B=ethyl acetate). Fractions containing product were pooled and concentrated to give C1 as a fine white powder. ¹H NMR (CDCl₃): 1.24-1.28 (m, 12H), 1.55-1.61 (m, 4H), 3.50 (t, 4H J=7.4 Hz), 6.68 (s, 4H). Measured mass=361.

Synthesis of C2 (Ex. 18)

Step 1. 3′ Hamino 5′ C6 disulfide siRNA (46.9 mg, 6.16 μmol) was dissolved in 9:1 DMSO/water (782 μl). TetraGalNAc (40.0 mg, 0.025 mmol) and DIEA (26.9 μl, 0.154 mmol) were dissolved in DMSO (200 μl), then added solution of HATU (14.0 mg, 0.037 mmol) in DMSO (141 μL) and stirred at RT for 15 minutes. This solution was added to the RNA solution and aged for 30 minutes. The reaction was diluted with DI water and dialyzed once to remove DMSO and purified by AEX (95:5-65:35 A:B linear gradient (A=20 mM TEA in 60% aqueous TFE; B=1M CsCl and 20 mM TEA in 60% aqueous TFE), Dionix Propac column). Fractions containing product were pooled, dialyzed, and lyophilized. Measured mass=9233.

Step 2. To this solid (30.8 mg, 3.34 μmol) was added TCEP (19.13 mg, 0.067 mmol) and DI water (2 mL). The reaction was stirred at RT for 1 hour, then aged overnight at 5° C. The reaction was diluted with DI water and dialyzed twice against DI water to give a solution of C2 that was used in further reactions without isolation.

Synthesis of C3 (Ex. 19)

C2 (60.1 mg, 6.60 umol, prepared in a manner analogous to B3) dissolved in DI water (37 mL) was treated with C1 (23.8 mg, 66.0 umol) dissolved in DMF (7 mL) to give a cloudy solution. The reaction was aged overnight, at which point dioxane (18 mL) was added to solubilize the reaction mixture. After aging for 30 additional minutes, the reaction was diluted with DI water. It was then dialyzed once against DI water, filtered, and purified by RP-HPLC (95:5-5:95% A:B linear gradient (A=100 mM aqueous TEAA; B=100 mM TEAA in acetonitrile) Waters Phenyl xbridge Column). Fractions containing product were dialyzed and lyophilized to give C3 as an amorphous white powder. Measured mass=9458.

Synthesis of C4-seq32 (Ex. 20)

C3 (10 mg, 1.057 umol) was dissolved in formamide modified with 20 mM MES buffer and 2 M thiourea (1 mL) and was added to P32 (8.62 mg, 2.11 umol). After 20 mins, LC-MS indicated good conversion to desired product. Reaction was purified by AEX (95:5-55:45 A:B linear gradient (A=20 mM TEA in 60% aqueous TFE; B=1M CsCl and 20 mM TEA in 60% aqueous TFE), Dionix Propac column). Fractions containing product were dialyzed to give C4-P32.

Synthesis of C6-seq32-(Ex. 21)

C4 (6.78 mg, 0.501 μmol) dissolved in DI water (3.40 mL) was treated with guide strand C5 (3.44 mg, 0.501 μmol) dissolved in DI water (530 μL). Analytical SAX indicated good duplex purity with some excess guide strand observed. Solution was lyophilized to give C6 as an amorphous white powder. Measured mass=passenger strand: 13539, guide strand: 6869.

Additional Synthesis of C6-Peptide Conjugates

Additional conjugates of C6 and Peptide Sequence were prepared in a manner analogous to that used for C6-seq32-c.

Examples 22-27 Preparation of C7 to C10, C11-P32 and C12-seq32-a

Scheme 8 as shown in FIG. 9A to FIG. 9E was used to prepare C7 to C10, C11-seq32 and C12-seq32.

Synthesis of C7 (Ex. 22)

Icosanedioic acid (600 mg, 1.752 mmol) was suspended in toluene (11 mL) and treated with DIEA (673 μL, 3.85 mmol) and DPPA (793 uL, 3.68 mmol). After stirring at room temp for 30 minutes, the reaction was slowly heated to 80° C., then to gentle reflux for two hours. Reaction was cooled and treated with tBuOH (1.675 mL, 17.52 mmol) and copper iodide (200 mg, 1.051 mmol) and heated back to reflux for 2 additional hours. Reaction was cooled (precipitation observed), diluted with DCM, filtered, and concentrated in vacuo. The crude product was purified by flash chromatography with a 100:0-0:50% A:B linear gradient (A=hexanes; B=ethyl acetate). Fractions containing product were pooled and concentrated to give C7. Measured mass=486.

Synthesis of C8 (Ex. 23)

C7 (101 mg, 0.208 mmol) was dissolved in DCM (20 mL) and treated with TFA (20 mL). The reaction was aged for five minutes, after which solvent and TFA were removed in vacuo to give C8 as a colorless oily solid that was used without further purification. Measured mass=286.

Synthesis of C9 (Ex. 24)

C8 (100.0 mg, 0.209 mmol) was suspended in chloroform (28 mL) and treated with tetrabutylammonium hydrogen sulfate (70.9 mg, 0.209 mmol), N-methoxy carbonyl maleimide (98.0 mg, 0.631 mmol), and DIEA (88.0 μL, 0.502 mmol). Saturated sodium bicarbonate (28 mL) was added. The reaction was stirred vigorously for 25 hours, after which it was extracted 3×50 mL DCM. The combined organic layers were dried with sodium sulfate, then evaporated to dryness. The crude product was purified by flash chromatography with a 100:0-0:50% A:B linear gradient (A=hexanes; B=ethyl acetate). Fractions containing the desired product were combined and evaporated to give C9. ¹H NMR (CDCl₃): 1.24-1.26 (m, 28H), 1.55-1.59 (m, 4H), 3.50 (t, 4H J=7.4 Hz), 6.68 (s, 4H). Measured mass=445.

Synthesis of C10 (Ex. 25)

C2 (12.0 mg, 1.31 μmol) was dissolved in 1:3 water:dioxane (14.4 mL) and was treated with C9 (5.8 mg, 13.1 μmol) dissolved in 1.4 mL dioxane. After aging overnight, the reaction was quenched with N-methyl maleimide (4.38 mg, 39.4 μmol) and was diluted with DI Water. The crude reaction was dialyzed once against DI water, filtered, and purified by RP-HPLC (95:5-5:95% A:B linear gradient (A=100 mM aqueous TEAA; B=100 mM TEAA in acetonitrile) Waters Phenyl xbridge Column). Fractions containing product were dialyzed against DI water and lyophilized to give C10. Measured mass: 9546.

Synthesis of C11-seq32 and C12-seq32-c (Ex. 26 and Ex. 27)

Conjugates C11-seq32 and C12-seq32-c were prepared in a manner analogous to that used for C4-seq32 and C6-seq32.

Additional Synthesis of C12-Peptide Conjugates

Additional conjugates of C12 and peptide sequence were prepared in a manner analogous to that used for C12-seq32.

Section D Examples 28-30 Preparation of C13, C14-seq32 and C15-seq32

Scheme 9 shown in FIG. 10A to FIG. 10D was used to prepare C13, C14-seq32 and C15-seq32-a.

Synthesis of C13 (Ex. 28)

C2 (11 mg, 1.22 μmol) dissolved in DI water (3.5 mL) was treated with C2 bismaleimide (2.69 mg, 12.20 umol) dissolved in DMF (270 μL). After one hour, LC-MS indicated good conversion to desired product. Reaction was dialyzed 3 times against DI water and lyophilized to give C13. Measured mass: 9317.

Synthesis of C14-seq32 (Ex. 29)

C13 (10.53 mg, 1.13 μmol) was dissolved in DI water (50 μL) and diluted with TFE modified with 50 mM AcOH (2.0 mL), then was added to seq32 (9.22 mg, 2.26 μmol) dissolved in 8M guanidine hydrochloride (60 μL). The reaction was aged for 10 minutes. Reaction was purified by AEX (95:5-55:45 A:B linear gradient (A=20 mM TEA in 60% aqueous TFE; B=1M CsCl and 20 mM TEA in 60% aqueous TFE), Dionix Propac column). Fractions containing product were dialyzed to give C14-seq32.

Synthesis of C15-seq32-c (Ex. 30)

C14-seq32 (9.81 mg, 0.738 μmol) dissolved in DI water (2.6 mL) was treated with guide strand C5 (7.76 mg, 0.738 μmol) dissolved in DI water (751 μL). Solution was lyophilized to give the desired product C15-seq32-c. Measured mass=passenger strand: 13396, guide strand: 6868

Additional Synthesis of C15-Peptide Conjugates

Additional conjugates of C15 and peptide sequence were prepared in a manner analogous to that used for C15-seq32.

Examples 31-33 Preparation of D1, D3 and D4

Scheme 10 as shown in FIG. 11A to FIG. 11 D was used to prepare D1, D3 and D4.

Synthesis of D1 (Ex. 31)

To a solution of NHS ester (100.0 mg, 0.320 mmol) in 0.5 mL anhydrous DCE were added azido amine (253.0 mg, 0.480 mmol) in 0.5 mL anhydrous DCE and 1.5 eq. triethylamine. The resulting solution was stirred for 1 h at room temperature, and the reaction mixture was loaded on a silica column, eluding with MeOH/DCM=0/100 to 10/90 over 25 min. The collected fraction was subject to LC-MS analysis and the result indicated >95% purity.

Synthesis of D3 (Ex. 32)

Oligonucleotide D2 (10 mg, 1.3 μmol) and azide linker D1 (5.6 mg, 7.8 μmol) were dissolved in degassed 3:1 DMA/water (1000 μL) in an Eppendorf tube, then a solution of copper(I) bromide-dimethyl sulfide (0.05 mg, 0.26 μmol) in degassed MeCN (100 μL) was added to the reaction mixture. After 60 min at 40° C., D2 was completely consumed monitored by LC-MS. The reaction mixture was diluted with 0.4 M EDTA (5 mL) and stirred for additional 15 min, then dialyzed against water using a Millipore 3K membrane and purified by RP HPLC (5%-60% A in B, A: 100 mM TEAA in MeCN, B: 100 mM TEAA in water). The product fractions were dialyzed against water and lyophilized to afford D3 as a white powder.

Synthesis of D4 (Ex. 33)

TetraGalNAc A10 (5.7 mg, 3.5 μmol), HATU (2.0 mg, 5.2 μmol), N,N-diisopropylethylamine (1.8 mg, 14 μmol) were dissolved in DMSO (100 μL). After 10 min, the activated ester was added to oligonucleotide D3 (6.4 mg, 0.70 μmol) in DMF (350 μL) and water (50 μL). The resulting reaction mixture was stirred for 15 min and quenched by addition of water, then purified by RP HPLC (5%-60% A in B, A: 100 mM TEAA in MeCN, B: 100 mM TEAA in water). The product fractions were dialyzed against water and lyophilized to afford R3 as a whiter powder.

Examples 34-35 Preparation of D5-Seq32 and D7-Seq32

Scheme 11 as shown in FIG. 12A-1 to FIG. 12B-2 was used to prepare D5-seq32 and D7-seq32.

Synthesis of D5-seq32 (Ex. 34)

Oligonucleotide D4 (6.5 mg, 0.60 μmol) in 200 μL formamide/pH=6.8 Tris buffer=3/1 was treated with peptide seq32 (9.8 mg, 2.4 μmol) in 200 μL of the same buffer and the resulting reaction mixture was stirred for 1 h. The reaction was diluted by addition of formamide 2.5 mL and purified by strong anion exchange chromatography on a Sepax Proteomix SAX NP10, 21.2×50 mm column (2%-30% B in A over 8 min, A: 60:40 trifluoroethanol:water, 40 mM triethylamine, B: 60:40 trifluoroethanol:water, 40 mM triethylamine, 1 M guanidine-HCl, 20 mL/min) to afford D5-seq32 as a white powder.

Synthesis of D7-seq32 (Ex. 35)

Oligonucleotide D5-seq32 (5.7 mg, 0.304 μmol) and the corresponding antisense strand D6 (2.0 mg, 0.29 μmol) were mixed in RNase free water for 1 h. The reaction mixture was lyophilized and the product D7-seq32-d was submitted for in vivo evaluation.

Synthesis of Additional D7-Peptide Conjugates

Additional conjugates of D7 and peptide sequence were prepared in a manner analogous to that used for D7-seq32.

Section E. Synthesis of Hybrid of Lipid and Peptide Conjugates Examples 36-42

Scheme 12 is shown in FIG. 13A to FIG. 13H-2.

Synthesis of E2 (Ex. 36)

Oligonucleotide E1 (300 mg, 39 μmol) and the PEG9 azide linker (58.5 mg, 78 μmol) were dissolved in degassed 3:1 DMA/water (10 mL) in a glass vial, then a solution of copper(I) bromide-dimethyl sulfide (20.06 mg, 98 μmol) in degassed DMSO (699 μL) was added to the reaction mixture. After 40 min at 45° C., E1 was completely consumed monitored by LC-MS. The reaction mixture was diluted with 0.4 M EDTA (20 mL) and stirred for additional 15 min, then dialyzed against water using a Millipore 3K membrane and lyophilized to afford E2 as a white powder.

Synthesis of E3 (Ex. 37)

TetraGalNAc A10 (237 mg, 145 μmol), HATU (55.2 mg, 145 μmol), N,N-diisopropylethylamine (94 mg, 726 μmol) were dissolved in DMSO (700 μL). After 10 min, the activated ester was added to oligonucleotide E2 (306 mg, 36 μmol) in DMA (7.5 mL) and water (2.5 mL). The resulting reaction mixture was stirred for 15 min and quenched by addition of water, then purified by RP HPLC (5%-60% A in B, A: 100 mM TEAA in MeCN, B: 100 mM TEAA in water). The product fractions were dialyzed against water and lyophilized to afford E3 as a whiter powder.

Synthesis of E4 (Ex. 38)

To a solution of E3 (246 mg, 24 μmol, 1 eq.) in water (8000 μL) was added TCEP-HCl (70 mg, 244 μmol, 10 eq.). The reaction mixture was mixed until TCEP-HCl fully dissolved. The solution was left at room temperature for 2 hours. The solution was centrifugal dialyzed two times against water over a 3K membrane to afford crude E4 which was directly used in the next step.

Synthesis of E5 (Ex. 39)

To a solution of E4 (244 mg, 24 μmol) in water (12 mL) was added N-(2-aminoethyl)maleimide trifluoroacetate salt (62.2 mg, 0.245 mmol, 10 eq.) dissolved in MeCN (0.5 mL). The solution was left at room temperature for 1 hour. LCMS indicated complete conversion. The solution was centrifugal dialyzed twice against water over a 3K membrane and lyophilized to afford E5 as a white powder.

Synthesis of E6 (Ex. 40)

E5 (40 mg, 3.95 μmol, 1 eq.) was dissolved in 4:1 DMA/water (500 μL). DIPEA (10.2 mg, 79 μmol, 20 eq.) was added to the above solution. Cholesterol chloroformate (18 mg, mol, 10 eq.) was dissolved in THF (500 μL). The two solutions were mixed together, and the reaction mixture was left at room temperature for 1 hour. LCMS indicated that the reaction was done. The reaction mixture was purified by RP HPLC (5%-95% B in A, A: 100 mM TEAA in water, B: 100 mM TEAA in MeCN). The product fractions were dialyzed against water and lyophilized to afford E6 as a whiter powder.

Synthesis of E7 (Ex. 41)

To a solution of E6 (24.5 mg, 2.3 μmol, 1 eq.) in water (1000 μL) was added piperidine in DMF (200 μL, 20% by volume, 200 eq.). The reaction mixture was left at room temperature for 1 hour. LCMS indicated that the reaction was done. The reaction mixture was filtered (0.2 uM), dialyzed against water, and lyophilized to give E7 as a whiter powder.

Synthesis of E8 (Ex. 42)

E7 (16 mg, 1.55 μmol, 1 eq.) was dissolved in freshly prepared aqueous sodium bicarbonate (0.1M, 400 μL). SPDP (4.85 mg, 0.016 mmol, 10 eq.) was dissolved in acetonitrile (400 uL). The two solutions were mixed together, and the reaction mixture was left at room temperature for 1 hour. The reaction mixture was purified by RP HPLC (5%-95% B in A, A: 100 mM TEAA in water, B: 100 mM TEAA in MeCN). The product fractions were dialyzed against water and lyophilized to afford E8 as a whiter powder.

Examples 43-44 Preparation of E8-Seq 137 and E9-Seq 137

Scheme 13 is shown in FIG. 14A-1 to FIG. 14B-2.

Synthesis of E9-Seq137 (Ex. 43)

Oligonucleotide E8 (3.0 mg, 0.286 μmol) in 100 μL of 2 M Thiourea/20 mM MES in Formamide pH 6.5 was treated with peptide seq 137 (2.33 mg, 0.572 μmol) in 100 μL of the same buffer and the resulting reaction mixture was left at RT for 30 min. The reaction was diluted by addition of formamide 1 mL and purified by strong anion exchange chromatography on a Propac SAX 22×250 mm column (5%-45% B in A over 15 min, A: 60:40 trifluoroethanol:water, 20 mM triethylamine, B: 60:40 trifluoroethanol:water, 20 mM triethylamine, 1 M guanidine-HCl, 20 mL/min) to afford E9-seq-137 as a white powder.

Synthesis of E10-Seq137-e (Ex. 44)

Passenger strand E9-seq137 (1.30 mg, 0.077 μmol) and the corresponding guide strand B7 (0.561 mg, 0.077 μmol) were mixed in RNase free water and heated to 90° C. for 1 min, then left at RT for 10 min. The duplex was lyophilized and the resulting product isolated as an amorphous white powder.

Synthesis of Additional E10-Peptide Conjugates

Additional conjugates of E10 and peptide sequence were prepared in a manner analogous to that used for E10-Seq137-e.

Section F. Preparation of 3, 13, 18 Tripeptide Conjugates Examples 45-49

Scheme 14 is shown in FIG. 15A to FIG. 15E-2.

Synthesis of Compound F2 (Ex. 45)

Compound A10 (210 mg, 0.129 mmol) was dissolved in dry N-methyl-2-pyrrolidinone (3 ml). HATU (48.9 mg, 0.129 mmol) and dry diisopropylethylamine (0.046 ml, 0.257 mmol) were added, and the mixture was sonicated until the solid was fully dissolved. The reaction was left at RT for 5 min. In a separate vial, compound F1 (500 mg, 0.0646 mmol) was dissolved in water (2 ml) and N-methyl-2-pyrrolidinone (5 ml). The A10 solution was added to the F1 solution, and the reaction was left at RT for 5 min. The reaction mixture was loaded on to an HPLC fitted with an Agilent PL-SAX 8 um 50×150 mm column heated to 60° C. The product was gradient eluted by starting at 100% solvent A (4:1 H₂O:ethanol, 20 mM triethylammonium acetate pH 7.0) and increasing to 80% solvent B (4:1 H₂O:ethanol, 20 mM triethylammonium acetate pH 7.0, 1M guanidinium hydrochloride) over 30 min at 100 ml/min. The fractions were combined, and the ethanol content was reduced to 5% by diluting with water. The solution was pump loaded onto a Waters XBridge 5 um 50×50 mm column at 50 ml/min, and the product was washed with water at 100 ml/min for 5 min. The desalted product was eluted by reversing the column and flowing 2:3 H₂O:acetonitrile at 50 ml/min through the column. The fraction was freeze dried to afford F2 as a white amorphous solid. Expected mass: 9363.6, found mass: 9363.5.

Synthesis of Compound F3 (Ex. 46)

F2 (500 mg, 0.0534 mmol) and azido-peg9-amine (253 mg, 0.481 mmol) were dissolved in 2,2,2-trifluoroethanol (5 ml) and water (5 ml). Nitrogen was bubbled through the solution for 1 min. In a separate vial, copper(I) bromide dimethyl sulfide (43.9 mg, 0.214 mmol) was dissolved in acetonitrile (2.5 ml). Nitrogen was bubbled through the solution for 1 min. The two solutions were mixed together, and nitrogen was bubbled through the reaction mixture for 1 min. The vial was sealed and left at RT for 1 hour. The reaction mixture was quenched with EDTA solution (0.5M, pH 8.0, 1 mL) and loaded onto an HPLC fitted with a Waters XBridge 5 um 50×250 mm column. The product was gradient eluted by starting at 100% solvent A (H₂O, 0.1M triethylammonium acetate pH 7.0) and increasing to 40% solvent B (acetonitrile) at 100 ml/min over 30 minutes. The fractions were combined, and the acetonitrile content was reduced to 5% by diluting with water. The solution was pump loaded onto a Waters XBridge 5 um 50×50 mm column at 50 ml/min, and the product was washed with water at 100 ml/min for 5 min. The desalted product was eluted by reversing the column and flowing 2:3 H₂O:acetonitrile at 50 ml/min through the column. The fraction was freeze dried to afford F3 as a white amorphous solid. Expected mass: 10943.5, found mass: 10943.2.

Synthesis of Compound F4 (Ex. 47)

F3 (467 mg, 0.0427 mmol) was dissolved in sodium bicarbonate solution (0.1M, 4.5 mL). NHS-SPDP (120 mg, 0.384 mmol) was dissolved in acetonitrile (1 mL). The solutions were mixed together, and the reaction was left at RT for 15 min. The reaction mixture was loaded onto an HPLC fitted with a Waters XBridge 5 um 50×250 mm column. The product was gradient eluted by starting at 100% solvent A (H₂O, 0.1M triethylammonium acetate pH 7.0) and increasing to 40% solvent B (acetonitrile) at 100 ml/min over 30 min. The fractions were combined, and the acetonitrile content was reduced to 5% by diluting with water. The solution was pump loaded onto a Waters XBridge 5 um 50×50 mm column at 50 ml/min, and the product was washed with water at 100 ml/min for 5 min. The desalted product was eluted by reversing the column and flowing 2:3 H₂O:acetonitrile at 50 ml/min through the column. The fraction was freeze dried to afford F4 as a white amorphous solid. Expected mass: 11535.3, found mass: 11535.1.

Synthesis of F5-Seq 463 (Ex. 48)

Peptide Seq. 612 (8.75 mg, 0.00520 mmol) was dissolved in DMSO (1 mL) containing 20 mM acetic acid. In a separate vial, F4 (10 mg, 0.000867 mmol) was dissolved in DMSO (1 ml) containing 20 mM acetic acid. The two solutions were mixed together and left at RT for 1 hour. The reaction was quenched with N-methylmaleimide (5.78 mg, 0.0520 mmol) and loaded onto an HPLC fitted with an Agilent PL-SAX 10 um 25×50 mm column. The product was gradient eluted by starting at 100% solvent A (2:3 H₂O:2,2,2-trifluoroethanol, 20 mM triethylamine) and increasing to 70% solvent B (2:3 H₂O:2,2,2-trifluoroethanol, 20 mM triethylamine, 0.5M guanidinium hydrochloride) at 30 ml/min over 20 min. The fractions were combined and loaded onto an HPLC fitted with a Waters XBridge 5 um 19×250 mm column. The product was gradient eluted by starting at 85% solvent A (H₂O, 0.1M hexylammonium acetate pH 7.0) and increasing to 65% solvent B (tetrahydrofuran) at 20 ml/min over 30 min. The fractions were combined, and the tetrahydrofuran content was reduced to less than 5% under vacuum. The solution was centrifugal dialyzed over a 10 k membrane once against water, once against 4:1 H₂O:ethanol containing 0.1M sodium chloride, and two more times against water. The concentrate was freeze dried to afford F5-Seq 463 as a white amorphous solid. Expected mass: 16247.8, found mass: 16247.9.

Example 49

Scheme 15 is shown in FIG. 16A-1 to FIG. 16B-2.

Synthesis of F6 Seq 463-f (Ex. 49)

F5-Seq 463 (7.75 mg, 0.000477 mmol) and Guide B7 (3.27 mg, 0.000477 mmol) were dissolved in H₂O (0.5 mL). The solution was left at RT for 1 hour and then freeze dried to afford the duplex of F6 Seq 463-f as a white amorphous solid (11 mg, quantitative). Expected mass of passenger strand: 16247.8, found mass: 16247.9. Expected mass of guide strand: 6852.5, found mass: 6852.7.

Synthesis of Additional F10-Peptide Conjugates an Duplexes

Additional conjugates of F10 and peptide sequences and their duplexes were prepared in a manner analogous to that used for F6-Seq 463-f.

Section G. Preparation of 3,8,13,18 Tetrapeptides Examples 50-53

Scheme 16 is shown in FIG. 17A-1 to FIG. 17D-2.

Synthesis of G2 (Ex. 50)

A10 (210 mg, 0.129 mmol) was dissolved in dry N-methyl-2-pyrrolidinone (3 ml). HATU (48.9 mg, 0.129 mmol) and dry diisopropylethylamine (0.046 ml, 0.257 mmol) were added, and the mixture was sonicated until the solid was fully dissolved. The reaction was left at RT for 5 min. In a separate vial, G1 (500 mg, 0.0643 mmol) was dissolved in water (2 ml) and N-methyl-2-pyrrolidinone (5 ml). The A10 solution was added to the G1 solution, and the reaction was left at RT for 5 min. The reaction mixture was loaded on to an HPLC fitted with an Agilent PL-SAX 8 um 50×150 mm column heated to 60° C. The product was gradient eluted by starting at 100% solvent A (4:1 H₂O:ethanol, 20 mM triethylammonium acetate pH 7.0) and increasing to 80% solvent B (4:1 H₂O:ethanol, 20 mM triethylammonium acetate pH 7.0, 1M guanidinium hydrochloride) over 30 minutes at 100 ml/min. The fractions were combined, and the ethanol content was reduced to 5% by diluting with water. The solution was pump loaded onto a Waters XBridge 5 um 50×50 mm column at 50 ml/min, and the product was washed with water at 100 ml/min for 5 min. The desalted product was eluted by reversing the column and flowing 2:3 H₂O:acetonitrile at 50 ml/min through the column. The fraction was freeze dried to afford the G2 as a white amorphous solid. Expected mass: 9399.7, found mass: 9399.5.

Synthesis of G3 (Ex. 51)

G2 (483 mg, 0.0514 mmol) and azido-peg9-amine (324 mg, 0.617 mmol) were dissolved in 2,2,2-trifluoroethanol (5 ml) and water (5 ml). Nitrogen was bubbled through the solution for 1 min. In a separate vial, copper(I) bromide dimethyl sulfide (50 mg, 0.244 mmol) was dissolved in acetonitrile (2.5 ml). Nitrogen was bubbled through the solution for 1 min. The two solutions were mixed together, and nitrogen was bubbled through the reaction mixture for 1 min. The vial was sealed and left at RT for 1 hour. The reaction mixture was quenched with EDTA solution (0.5M, pH 8.0, 1 mL) and loaded onto an HPLC fitted with a Waters XBridge 5 um 50×250 mm column. The product was gradient eluted by starting at 100% solvent A (H₂O, 0.1M triethylammonium acetate pH 7.0) and increasing to 40% solvent B (acetonitrile) at 100 ml/min over 30 min. The fractions were combined, and the acetonitrile content was reduced to 5% by diluting with water. The solution was pump loaded onto a Waters XBridge 5 um 50×50 mm column at 50 ml/min, and the product was washed with water at 100 ml/min for 5 min. The desalted product was eluted by reversing the column and flowing 2:3 H₂O:acetonitrile at 50 ml/min through the column. The fraction was freeze dried to afford G3 as a white amorphous solid. Expected mass: 11506.2, found mass: 11506.0.

Synthesis of G4 (Ex. 52)

G3 (455 mg, 0.0396 mmol) was dissolved in sodium bicarbonate solution (0.1M, 5 mL). NHS-SPDP (160 mg, 0.512 mmol) was dissolved in acetonitrile (1.5 mL). The solutions were mixed together, and the reaction was left at RT for 15 min. The reaction mixture was loaded onto an HPLC fitted with a Waters XBridge 5 um 50×250 mm column. The product was gradient eluted by starting at 100% solvent A (H₂O, 0.1M triethylammonium acetate pH 7.0) and increasing to 40% solvent B (acetonitrile) at 100 ml/min over 30 min. The fractions were combined, and the acetonitrile content was reduced to 5% by diluting with water. The solution was pump loaded onto a Waters XBridge 5 um 50×50 mm column at 50 ml/min, and the product was washed with water at 100 ml/min for 5 min. The desalted product was eluted by reversing the column and flowing 2:3 H₂O:acetonitrile at 50 ml/min through the column. The fraction was freeze dried to afford G4 as a white amorphous solid. Expected mass: 12295.3, found mass: 12295.1.

Synthesis of G5-Seq 489 (Ex. 53)

Peptide SEQ ID NO: 489 (CIFGAIAGFIKNIWEGLI all (D)) (13.6 mg, 0.00694 mmol) was dissolved in DMSO (1 mL) containing 20 mM acetic acid. In a separate vial, G4 (10 mg, 0.000867 mmol) was dissolved in DMSO (1 ml) containing 20 mM acetic acid. The two solutions were mixed together and left at RT for 1 hour. The reaction was quenched with N-methylmaleimide (7.71 mg, 0.0694 mmol) and loaded onto an HPLC fitted with an Agilent PL-SAX 10 um 25×50 mm column. The product was gradient eluted by starting at 100% solvent A (2:3 H₂O:2,2,2-trifluoroethanol, 20 mM triethylamine) and increasing to 70% solvent B (2:3 H₂O:2,2,2-trifluoroethanol, 20 mM triethylamine, 0.5M guanidinium hydrochloride) at 30 ml/min over 20 min. The fractions were combined and loaded onto an HPLC fitted with a Waters XBridge 5 um 19×250 mm column. The product was gradient eluted by starting at 85% solvent A (H₂O, 0.1M hexylammonium acetate pH 7.0) and increasing to 65% solvent B (tetrahydrofuran) at 20 ml/min over 30 min. The fractions were combined, and the tetrahydrofuran content was reduced to less than 5% under vacuum. The solution was centrifugal dialyzed over a 10 k membrane once against water, once against 4:1 H₂O:ethanol containing 0.1M sodium chloride, and two more times against water. The concentrate was freeze dried to afford G5-Seq 489 as a white amorphous solid. Expected mass: 19708.1, found mass: 19708.0.

Example 54

Scheme 17 is shown in FIG. 18A-1 to FIG. 18B-2.

Synthesis of G6-Seq 489-g (Ex. 54)

G5-Seq 489 (8.5 mg, 0.000434 mmol) and B7 (2.98 mg, 0.000434 mmol) were dissolved in H₂O (0.5 mL). The solution was left at RT for 1 hour and then freeze dried to afford the duplex G6-Seq 489-g as a white amorphous solid. Expected mass of passenger strand: 19708.1, found mass: 19708.3. Expected mass of guide strand: 6852.5, found mass: 6852.6.

Synthesis of Additional G6-peptide Conjugates and Duplexes

Additional conjugates of G6 and peptide sequences and their duplexes were prepared in a manner analogous to that used for G6-Seq 489-g.

SECTION H. Preparation of 3,8,13,18 tetrapeptide Examples 55-58

Scheme 18 below was used to prepare H1 to H5.

Synthesis of H1 (Ex. 55)

Into a 500-mL round-bottom flask purged and maintained with an inert atmosphere of nitrogen was placed a solution of di-tert-butyl 1-(tert-butylthio)hydrazine-1,2-dicarboxylate (15 g, 46.8 mmol, 2.00 equiv) in N,N-dimethylformamide (30 mL). A solution of 2-aminoethanethiol hydrochloride (2.66 g, 23.4 mmol, 1 eqiv) in N, N-dimethylformamide (80 ml) was added slowly into the round-bottom flask. This was followed by the addition of triethylamine (2.36 g, 23.4 mmol, 1 equiv). After stirring at RT overnight, a white solid was precipitating. Dry N, N-dimethylformamide (100 ml) was added to obtain a nearly clear solution. Triethylamine was added until a white solid was precipitating again. The reaction mixture was stirred at RT for 8 hours. The solution was filtered and evaporated under reduced pressure. Diethyl ether (200 ml) was added to the residue and filtered. The white solid was collected and dried in dessicator. Afterward, this white solid was dissolved five times in diethyl ether (5×10 ml), stirred for several minutes and filtered. The desired product was obtained as a white solid. ¹HNMR (CDCl₃, 500 MHz, ppm): 1.36 (s, 9H), 3.07 (t, 2H), 3.4 (t, 2H), 8.3 (s, 2H).

Synthesis of H3 (Ex. 56)

Lithocholic acid (H2) (7 gm, 18.59 mmol, 1 equiv) was dissolved in dry dicholormethane (200 ml) and then cooled to 0° C. Following this N, N-dicyclohexylcarbodiimide (4.6 g, 22.31 mmol, 1.2 equiv) was added to the solution. After stirring for 30 min at 0° C., pentafluorophenol (3.76 gm, 20.45 mmol, 1.1 equiv) in dichloromethane (13 ml) was added. Stirring was then continued at RT under argon for an additional 20 h. The precipitated N, N-dicyclohexylurea was filtered off and washed with cold dichloromethane. Combined filterates were then evaporated under reduced pressure. The oily residue obtained was then diluted with dichloromethane (50 ml) and washed with sat. aq. NaCl (60 ml) and water (80 ml). The organic phase was dried over Na₂SO₄, filtered and evaporated to dryness. The dried compound was purified using column chromatography (elution with CH₂Cl₂/CH₃OH, 100/0-97/3). MS (m/z); 566 [M+Na]⁺

Synthesis of H4 (Ex. 57)

Compound H3 (4.5 gm, 8.29 mmol, 1 equiv) was dissolved in dry dichloromethane (15 ml) and then cooled to 0° C. A cold mixture of 2-(tert-butyldisulfanyl)ethanamine (H1) (2.057 gm, 12.44 mmol, 1.5 equiv) and triethylamine (2.56 gm, 2.52 mmol, 3 equiv) in dichloromethane (7 ml) was added to the resulting solution. The reaction mixture was stirred at RT for 2 h. TLC confirmed the formation of product. The reaction mixture was washed with sat. aq. NaCl (20 ml×2) and water (20 ml×2). The organic phase was dried over Na₂SO₄, filtered and dried over vacuum. The crude product was purified via silica gel column chromatography (elution with CH₂Cl₂/CH₃OH, 100/0-95/5) yielding pure compound H4. MS (m/z); 524.35, [M+1]⁺

Synthesis of H5 (Ex. 58)

H4 (3 gm, 5.73 mmol, 1 equiv) was dissolved in dry dichloromethane (15 ml) and triethylamine was added (0.869 g, 8.59 mmol, 1.5 equiv). The reaction mixture was cooled to 0° C. 2-Cyanoethyl-N, N-diisopropylaminochlorophosphite (2.71 gm, 11.45 mmol, 2 equiv) in dry dichloromethane (10 ml) was added dropwise to the reaction mixture. The resulting solution was stirred for 1 h. TLC confirmed the formation of product. The reaction mixture was evaporated and purified on silica gel column (elution with hexanes/ethylacetate/triethylamine, 100/0/1.5 to 60/40/1.5). MS (m/z); 724.46 [M+1]⁺³¹P NMR (CDCl₃, 500 MHz, ppm); 146.5

Examples 59-66

Scheme 19 as shown in FIG. 19A to FIG. 19I-2 was used to prepare Ex. 59 to Ex. 66.

Synthesis of H6 (Ex. 59)

See synthesis of B2 for reaction procedure. Expected mass: 9609.071, found mass: 9605.

Synthesis of H7 (Ex. 60)

To a solution of H6 (15 mg, 1.56 umol, 1 eq) in water (1400 ul) was added TCEP-HCl (26.8 mg, 0.094 mmol, 60 eq). The reaction mixture was mixed until TCEP-HCl fully dissolved. The solution was left at RT overnight. The solution was centrifugal dialyzed two times against water over 3K membrane. Expected mass: 9520, found mass: 9517.

Synthesis of H8 (Ex. 61)

See synthesis of B9 for reaction procedure. Expected mass: 9630, found mass: 9627.

Synthesis of H9-Seq32 (Ex. 62)

See the synthesis of B10-seq32 for reaction procedure. Expected mass: 13597, found mass: 13598.

Synthesis of H7-Seq32-h (Ex. 63)

See the synthesis of B11-seq32 for reaction procedure.

Synthesis of H8 (Ex. 64)

See the synthesis of C13 for reaction procedure. Expected mass: 9741.

Synthesis of H9-Seq32 (Ex. 65)

See the synthesis of C14 for reaction procedure. Expected mass: 13819, found mass: 13820.

Synthesis of H10-Seq32-h (Ex. 66)

See the synthesis of C15-Seq32 for reaction procedure.

Additional Synthesis of H7 and H10 Peptide Conjugates

Additional conjugates of H7 and H10 and peptide sequences and their duplexes were prepared in a manner analogous to that used for H7-Seq32-h and H10-Seq32-h.

Section I. Preparation of 3,13,18 Trienzymatic Cleavable Linker Peptide Conjugates Examples 67-73

Scheme 20 is shown in FIG. 20A-1 to FIG. 20E-2.

Synthesis of 13 (Ex. 67)

I1 (160 mg, 0.209 mmol) and I2 (48.8 mg, 0.219 mmol) were dissolved in DMA (1 mL) and were treated with N-methylmorpholine (46 μL, 0.417 mmol). The reaction was stirred at RT for 6 hours, then purified by RP-HPLC (95:5-20:80% A:B linear gradient (A=0.1% aqueous TFA; B=0.1% TFA in acetonitrile) Waters C18 xbridge Column 19×250 mm). Fractions containing I3 were extracted with 2:1 DCM:MeOH, dried over Na₂SO₄, filtered, and concentrated in vacuo to give the product. Measured mass=814.3

Synthesis of I4 (Ex. 68)

I3 (88 mg, 0.108 mmol) was dissolved in DMA (1 mL) and was treated with piperidine (200 μL, 2.02 mmol) and stirred at 10° C. for 10 min. TFA (156 μL, 2.02 mmol) was added to quench the reaction. The reaction mixture was purified by RP-HPLC (95:5-60:40% A:B linear gradient (A=0.1% aqueous TFA; B=0.1% TFA in acetonitrile) Waters C18 xbridge Column 30×250 mm). Fractions containing 14 were lyophilized to give the product. Measured mass=592.3.

Synthesis of I5 (Ex. 69)

I4 (912 mg, 1.324 mmol) was dissolved in DMSO (7.7 mL) and treated with L1 (1.0 g, 1.40 mmol) and DIEA (463 μL, 2.65 mmol). The reaction mixture was stirred for 15 min and was purified by RP-HPLC (100:0-0:100% A:B linear gradient (A=0.1% aqueous TFA; B=0.1% TFA in acetonitrile) Waters C18 xbridge column. Fractions containing I5 were lyophilized to give the product. Measured mass=609.5 [M+2]

Synthesis of I7 (Ex. 70)

I6 (500 mg, 0.065 mmol) and I5 (236 mg, 0.194 mmol) were dissolved in a pH 5.5 MES buffer (51.6 ml, 500 mM) and acetonitrile (12.91 ml). The solution was degassed with nitrogen for 10 min, after which it was treated with CuBr.SMe₂ (133 mg, 0.646 mmol) and degassed for an additional five minutes with nitrogen. The reaction mixture was sonicated and stirred for 30 min, then purified by RP-HPLC (95:5-5:95% A:B linear gradient (A=100 mM aqueous TEAA; B=100 mM TEAA in acetonitrile) Waters Phenyl xbridge Column). Fractions containing product were dialyzed twice against 0.32M EDTA pH 6.5 over a 3K membrane, then three times against water. The concentrate was then dialyzed twice against 200 mM TEAA and then three times against water. The concentrate was lyophilized to give the product as an amorphous white solid. Measured mass=11400

Synthesis of I8 (Ex. 71)

I7 (287 mg, 0.025 mmol) was suspended in water (100 uL) and diluted with NMP (2.0 mL), which produced a homogeneous solution upon standing. HATU (13 mg, 0.035 mmol) was dissolved in NMP (200 uL) and was added to A10 (62 mg, 0.038 mmol). The reaction mixture was diluted with NMP (200 uL) and was then treated with DIEA (13 uL, 0.076 mmol). The HATU reaction mixture was then added to the RNA solution in one portion and aged for 10 min. Reaction was diluted with DI water and purified by RP-HPLC (95:5-5:95% A:B linear gradient (A=100 mM aqueous TEAA; B=100 mM TEAA in acetonitrile) Waters Phenyl xbridge Column). Fractions containing 18 were dialyzed three times against water over a 3K membrane. The concentrate was lyophilized to give the product as an amorphous white solid. Measured mass=13027.

Synthesis of I9-Seq 1681 (Ex. 72)

I8 (20 mg, 1.537 μmol) was dissolved in TFE modified with 50 mM AcOH (2 mL). In a separate vial, Seq ID 1681 (8.63 mg, 6.15 umol) was suspended in 8M Gn.HCl (400 uL) and was diluted with 50 mM AcOH in TFE (2 mL) to form a slightly cloudy suspension, then added to the RNA solution. After 10 min, more Seq ID 1681 (8.63 mg, 1.54 umol) was added and the reaction was aged 30 min, after which AEX indicated near-complete conversion to desired product. Reaction was quenched with N-methylmaleimide (6.83 mg, 61.5 μmol) and was purified by AEX (0-40% 1M Gn.HCl in 1:1 water:TFE with 40 mM TEAA pH 7.5, Proteomix NP10 column heated to 60° C.). Material was repurified using 70:30-25:75 gradient of 200 mM HAA pH 7.5:ACN and an Agilent PLRP-S column. Pure fractions were pooled, dialyzed, and lyophilized to give 19-Seq 1681 (6.37 mg, 0.302 μmol, 19.65% yield).

Synthesis of I10-Seq 1681-f (Ex. 73)

I9-seq 1681 (3.02 mg, 0.143 μmol) was dissolved in water (950 μl) and was treated with a solution of B7 (0.980 mg, 0.143 μmol) in water (144 μl). The reaction mixture aged for 15 min and was then lyophilized to give the product as an amorphous white solid. Measured mass=21107.

Additional Synthesis of I10 Peptide Conjugates an Duplexes

Additional conjugates of I10 and peptide sequences and their duplexes were prepared in a manner analogous to that used for I10-seq-1681-f.

Section J. Preparation of Amino Modified C2 Linkers Examples 74-82

Scheme 21 is shown in FIG. 21A to FIG. 21H-2.

Synthesis of A10B (Ex. 74)

In a test tube equipted with a stir bar, A10 (100 mg, 0.061 mmol) was dissolved in DMSO (611 μl) followed by the addition of Hunig's Base (133 μl, 0.764 mmol) and HATU (76 mg, 0.199 mmol). After 20 min, N-(2-aminoethyl)maleimide trifluoroacetate salt (12.85 mg, 0.092 mmol) dissolved in 400 μL of DMSO was added. After 20 min, the reaction was determined complete and quenched with water (1.5 mL) until yellow color almost dissipated. The reaction was purified by reverse phase chromatography (Gilson 2020, Solvent A) 0.1% TFA in water/Solvent B) 0.1% TFA in ACN, 0-50% gradient for 15 min, 40 mL/min, XBridge Prep C18 5 μm OBD 30×250 mm). The resulting fractions were lyophilized to afford a white solid, A10B. [M+1, expected]=1757.807, [M+1, observed]=1759.0.

Synthesis of J2 (Ex. 75)

See Synthesis of B3 for reaction procedure. J2 [M+1, expected]=7604.750, [M+1, observed]=7600.0.

Synthesis of J3 (Ex. 76)

A10B (10.26 mg, 5.84 μmol) was dissolved in water (700 μL) and added to a 1.8 mL solution (1 water: 1 acetate buffer: 2 formamide) of J2 (29.6 mg, 3.89 μmol). The reaction was shaken at RT for 20 min and then determined complete. The reaction mixture was purified using strong anion exchange chromatography (Gilson PLC 2020, Sepax Proteomix SAX NP10 21.2×50 mm, Buffer A: 3:2 trifluoroethanol:water, 40 mM triethylamine/Buffer B: 3:2 trifluoroethanol:water, 40 mM triethylamine, 1000 mM guanidine-HCl, 1% B hold for 3 minutes, then 5% B-45% B over 12 minutes). The fractions were dialyzed three times against water over a 3K membrane to afford a white solid, J3. [M+1, expected]=9362.556, [M+1, observed]=9359.0.

Synthesis of J4 (Ex. 77)

To an Eppendorf vial, J3 (6.34 mg, 0.678 μmol) was dissolved in water (250 μL). In a separate Eppendorf vial, N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) (0.831 mg, 2.035 μmol) was dissolved in DMSO (50 μL). The SPDP solution was added to the RNA solution. After 4 hours, the reaction was recharged with additional SPDP (2.77 mg, 6.78 μmol) which was dissolved in 50 μL DMSO. After 24 hr, the reaction was recharged with additional SPDP (2.77 mg, 6.78 μmol) which was dissolved in 50 μL DMSO. After 72 hr, the reaction was diluted to 3 mg/mL with the addition of 390 μL of pH 8.1 sodium bicarbonate. After 2 hr, an additional 3eq. of SPDP in 50 μL DMSO were added. The reaction mixture was dialyzed three times against water over a 3K membrane and lyophilized to afford a white solid, J4. [M+1, expected]=9543.834, [M+1, observed]=9554.0.

Synthesis of J5-Seq26 (Ex. 78)

See Synthesis of B10-Seq32 for reaction procedure. J5-Seq26-Mass observed: 11413.

Synthesis of J6-Seq26-i (Ex. 79)

See Synthesis of B11-Seq32-b for reaction procedure. J6-Seq26-i—Mass observed: 18265.

Synthesis of J7 (Ex. 80)

To an Eppendorf vial, J3 (5.8 mg, 0.621 μmol) was dissolved in water (250 μL). In a separate Eppendorf vial, Succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) (0.727 mg, 1.862 μmol was dissolved in DMSO (50 μL) and the pH was adjusted to pH 5 with the addition of 1 small drop of TFA. The SMCC solution was added to the RNA solution. After several hours, the pH was titrated to pH 7 with the gradual addition of 0.1N NaOH. After 18 hr, 6 eq. of SMCC were dissolved in 50 μL DMSO and added to the reaction mixture. After 4 hr, an additional 3eq. of SMCC in 50 μL DMSO was added to the reaction. After several hr, 300 μL of pH 8.1 sodium bicarbonate solution was added to the reaction. The reaction was dialyzed three times against water over a 3K membrane and lyophilized to afford a white solid, J7. [M+1, expected]=9543.834, [M+1, observed]=9554.0.

Synthesis of J8-Seq26 (Ex. 81)

See Synthesis of B10-Seq32 for reaction procedure. J8-Seq26-Mass observed: 11545.

Synthesis of J9-Seq26-i (Ex. 82)

See Synthesis of B11-Seq32 for reaction procedure. J9-Seq26-I—Mass expected: 18397.

Additional Synthesis of J6 & J9 Peptide Conjugates

Additional conjugates of J6 and J9 and peptide sequences and their duplexes were prepared in a manner analogous to that used for J6-Seq26, J9-Seq26 and J6-Seq26-i, J9-Seq26-i.

Section K. 3′ Bis Peptide Linkers Examples 83-87

Scheme 22 is shown in FIG. 22A-1 to FIG. 22D-2.

Synthesis of K2 (Ex. 83)

In a 20 mL vial, 3-(tritylthio)propanoic acid (158 mg, 0.454 mmol) was dissolved in DMF (1.514 mL) followed by the addition of HATU (184 mg, 0.484 mmol) and Hunig's base (0.158 mL, 0.908 mmol). The reaction solution turned light yellow in color. After 5 min, K1 (100 mg, 0.151 mmol) was added as a solid and the reaction solution turned transparent orange in color. The reaction was stirred at RT for 15 min and then determined complete.

The reaction was purified by reverse phase chromatography (Gilson 2020, 5-95% ACN/Water with a 0.1% TFA modifier, flow rate: 20 mL/min, gradient time: 22 min, column: XBridge prep OBD 5 m C18 19×250 nm). The resulting fractions were lyophilized to afford a white solid, K2. [M+1, expected]=877.059, [M+1, observed]=877.4

Synthesis of K3 (Ex. 84)

In an Eppendorf vial, K2 (10.07 mg, 0.011 mmol) was dissolved in formamide (0.5 mL). In a 15 mL Falcon tube, peptide Seq ID 74 (57.92 mg, 0.034 mmol) was dissolved in formamide (1 mL). The peptide/formamide solution was added to the linker/formamide solution and stirred at RT for 20 min.

The reaction was determined complete and the reaction was purified by reverse phase chromatography (Gilson 2020, 5-100% ACN/Water with a 0.1% TFA modifier, flow rate: 20 mL/min, gradient time: 30 minutes, column: XBridge prep OBD 5 m C18 19×250 nm). The resulting fractions were lyophilized to afford a white solid, K3. [M+3, expected]=1416.03, [M+3, observed]=1415.0

Synthesis of K4 (Ex. 85)

In a 40 mL vial, a solution of TFA (1000 μL), water (96 μL), and triisopropylsilane (96 μL) in a 0.83:0.08:0.08 mixture by volume was combined and added to K3 (47 mg, 0.011 mmol) in a 20 mL vial which was stirred at RT for 10 min. An additional 500 μL of TFA was added and the reaction was stirred for an additional 10 min. The reaction was determined complete, concentrated under reduced pressure, diluted with 3.5 mL of 2M thiourea pH 6.5 in FMD and MES, and purified by reverse phase chromatography (Gilson 2020, 5-80% ACN/Water with a 0.1% TFA modifier, flow rate: 20 mL/min, gradient time: 20 minutes, column: XBridge prep OBD 5 μm C18 19×250 nm). The resulting fractions were lyophilized to afford a white solid, K4. [M+3, expected]=1334.34, [M+3, observed]=1334.4

Synthesis of K5-Seq 74 (Ex. 86)

See Synthesis of B10-Seq32 for reaction procedure. K5-Seq 74-Expected mass: 13178.103.

Synthesis of K6-Seq 74-b (Ex. 87)

See Synthesis of B10-Seq32 for reaction procedure. Observed mass passenger=15907; Observed mass guide=8744; duplex=24651.

Additional Synthesis of KS Peptide Conjugates and Duplexes

Additional conjugates of KS and peptide sequences and the corresponding duplexes were prepared in a manner analogous to that used for K5-Seq 74 and K6-Seq 74-b.

Section L. Preparation of Guide Strand Position 2′-10,15 ECL Peptide Conjugates Examples 88-94

Scheme 23 is shown below, and in FIG. 23A to FIG. 23C-2.

Synthesis of L3 (Ex. 88)

(9H-fluoren-9-yl)methyl ((S)-3-methyl-1-(((S)-1-((4-((((4-nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxo-5-ureidopentan-2-yl)amino)-1-oxobutan-2-yl)carbamate L1 (500 mg, 0.652 mmol), 2-(pyridin-2-yldisulfanyl)ethanamine hydrochloride (153 mg, 0.685 mmol), and N-methylmorpholine (0.143 mL, 1.30 mmol) were dissolved in N,N-Dimethylacetamide (3 mL). The reaction mixture was aged for 16 h at RT and purified by reverse phase chromatography on a Waters Xbridge C18 column (5 uM, 30×250 mm) using a gradient of 5-80% ACN/water with 0.1% TFA over 20 min at 40 mL/min. The product was lyophilized to give L3 as a solid. MS (m/z): 814 (M+1).

Synthesis of L4 (Ex. 89)

(9H-fluoren-9-yl)methyl ((S)-3-methyl-1-oxo-1-(((S)-1-oxo-1-((4-((((2-(pyridin-2-yldisulfanyl)ethyl)carbamoyl)oxy)methyl)phenyl)amino)-5-ureidopentan-2-yl)amino)butan-2-yl)carbamate L3 (343 mg, 0.421 mmol) and piperidine (200 uL, 2.02 mmol) were dissolved in N,N-Dimethylacetamide (3 mL). The reaction mixture was aged for 10 min at RT, quenched with trifluoroacetic acid (156 uL, 2.02 mmol), and purified by reverse phase chromatography on a Waters Xbridge C18 column (5 uM, 30×250 mm) using a gradient of 5-40% acetonitrile/water with 0.1% trifluoroacetic acid over 20 min at 40 mL/min. The product was lyophilized to give L4 as a solid. MS (m/z): 592 (M+1).

Synthesis of L6 (Ex. 90)

To a solution of 4-((S)-2-((S)-2-amino-3-methylbutanamido)-5-ureidopentanamido)benzyl (2-(pyridin-2-yldisulfanyl)ethyl)carbamate L4 (238 mg, 0.346 mmol) in dimethylsulfoxide (1.5 mL) was added a solution of bis(2,5-dioxopyrrolidin-1-yl) octanedioate L5 (509 mg, 1.382 mmol) and triethylamine (0.096 mL, 0.691 mmol). The reaction mixture was aged for 15 min and purified on a silica gel column (80 g) using a gradient of 1-10% methanol/dichloromethane over 30 min at 60 mL/min to give L6 as a solid. MS (m/z): 845 (M+1)

Synthesis of L8 (Ex. 91)

RNA compound L7 (163 mg, 0.024 mmol) and 2-azidoethanamine hydrochloride (30 mg, 0.245 mmol) were dissolved in an argon degassed, 3:1 mixture of N,N-Dimethylacetamide:water (2 mL). An argon degassed solution of copper (I) bromide dimethyl sulfide complex (12 mg, 0.059 mmol) was added and the mixture was aged at 45° C. for 16 h. The mixture was quenched with a 0.5 M solution of EDTA (3 mL) and let stand for 15 min. The product was isolated by spin dialysis against water (3×) followed by lyophilization to give a solid. MS (m/z): 7086.

Synthesis of L9 (Ex. 92)

RNA compound L8 (46 mg, 6.49 μmol) and N-methylmorpholine (7.1 mL, 65 μmol) were dissolved in water (250 μL) and DMSO (250 μL) at 10° C. To this mixture was added a solution of 2, 5-dioxopyrrolidin-1-yl 8-(((S)-3-methyl-1-oxo-1-(((S)-1-oxo-1-((4-((((2-(pyridin-2-yldisulfanyl)ethyl)carbamoyl)oxy)methyl)phenyl)amino)-5-ureidopentan-2-yl)amino)butan-2-yl)amino)-8-oxooctanoate L6 (18 mg, 21 μmol) dissolved in DMSO (500 μL). The reaction mixture was aged for 16 h, diluted with water (1.5 mL) and purified by ion pairing chromatography on a Waters Xbridge phenyl column (5 μM, 19×250 mm) using a gradient of 0-55% acetonitrile/water with 100 mM triethylammonium acetate over 15 min at 20 mL/min. The product was isolated by spin dialysis against water (3×) followed by lyophilization to give a solid. MS (m/z): 8547.

Synthesis of L10-Seq 463 (Ex. 93)

RNA compound L9 (11 mg, 1.29 μmol) was dissolved in trifluoroethanol containing 50 mM acetic acid (500 μL). To this solution was added peptide Seq 463 (8.66 mg, 5.15 μmol) dissolved in trifluoroethanol containing 50 mM acetic acid (1000 μL). The mixture was aged for 10 min, quenched with N-methylmaleimide (1.9 mg, 44 μmol), and purified by ion pairing chromatography on a Waters Xbridge phenyl column (10 μM, 19×250 mm) using a gradient of 5-95% acetonitrile/water with 100 mM triethylammonium acetate over 15 min at 20 mL/min. The product was isolated by spin dialysis against water (3×) followed by lyophilization to give a solid. MS (m/z): 11687.

Synthesis of L11-Seq 463-i (Ex. 94)

A solution of L10-Seq 463 (2.46 mg, 0.27 μmol) dissolved in DI water (300 μL) was added to B2 (3.1 mg, 0.27 μmol) and heated at 90° C. for 1 min. Solution was lyophilized to give duplex as a whilte solid. MS (m/z) passenger strand: 9267, guide strand: 11686.

Additional Synthesis of L10 Peptide Conjugates and L11 Duplexes

Additional L10 conjugates of peptide sequences and the corresponding duplexes L11 were prepared in a manner analogous to that detailed above.

Section M. Synthesis of Guide Strand Position 2′-10,15 Disulfide Peptide Conjugates Examples 95-98

Scheme 24 is shown in FIG. 24A-1 to FIG. 24B-2.

Synthesis of M1 (Ex. 95)

3-(Pyridin-2-yldisulfanyl)propanoic acid (506 mg, 2.35 mmol), 2-azidoethanamine hydrochloride (317 mg, 2.59 mmol), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (496 mg, 2.59 mmol), 1-hydroxy-7-azabenzotriazole (199 mg, 1.46 mmol), and n-methylmorpholine (0.44 mL, 4.7 mmol) were dissolved in dichloromethane (25 mL). The mixture was aged for 1 h, diluted with saturated sodium bicarbonate solution (25 mL) and organic layer separated. Extracted aqueous later with dichloromethane (2×25 mL), dried combined organics over anhydrous sodium sulfate, filtered off solids and concentrated in vacuo. The mixture was purified on a silica gel column (80 g) using a gradient of 0-50% ethyl acetate/dichloromethane over 15 min at 30 mL/min to give a clear oil of M1. MS (m/z): 284.

Synthesis of M2 (Ex. 96)

RNA compound L7 (180 mg, 26 μmol) and M1 (59 mg, 208 μmol) were dissolved in a 100 mM, pH 5.5 MES buffer (3.6 mL) and acetonitrile (0.9 mL). This mixture was degassed with argon for 15 min. To this solution was added a degassed solution of copper (I) bromide dimethyl sulfide complex (13 mg, 65 μmol) dissolved in acetonitrile (0.45 mL) and aged at RT for 28 h. The mixture was quenched with a 100 mM, pH 8 solution of EDTA (5 mL) and allowed to stand for 15 min. The mixture was purified by ion pairing chromatography on a Waters Xbridge phenyl column (5 μM, 30×150 mm) using a gradient of 0-30% acetonitrile/water with 100 mM triethylammonium acetate over 15 min at 30 mL/min. The product was isolated by spin dialysis against water (3×) followed by lyophilization to give a solid of M2. MS (m/z): 7481.

Synthesis of M3-Seq 463 (Ex. 97)

RNA compound M2 (27.3 mg, 3.65 μmol) was dissolved in trifluoroethanol containing 50 mM acetic acid (1300 μL). To this solution was added peptide Seq 463 (15.4 mg, 9.13 μmol) dissolved in trifluoroethanol containing 50 mM acetic acid (1300 μL). The mixture was aged for 10 min, quenched with N-methylmaleimide (10.1 mg, 91 μmol), and purified by ion pairing chromatography on a Waters Xbridge phenyl column (10 μM, 19×250 mm) using a gradient of 5-80% acetonitrile/water with 100 mM triethylammonium acetate over 15 min at 20 mL/min. The product was isolated by spin dialysis against water (3×) followed by lyophilization to give a solid of M3-Seq 463. MS (m/z): 10624.

Synthesis of M4-Seq 463-j (Ex. 98)

A solution of B2 (2.18 mg, 0.24 μmol) dissolved in DI water (290 μL) was added to M3-Seq 463 (2.5 mg, 0.24 μmol) and heated at 90° C. for 1 min. This solution was lyophilized to give duplex M4-Seq 463-j as a whilte solid. MS (m/z) passenger strand: 9267, guide strand: 10621

Additional Synthesis of M3 Peptide Conjugates and M4 Duplexes

Additional M3 conjugates of peptide sequences and the corresponding duplexes M4 were prepared in a manner analogous to that detailed above.

Section N. Synthesis of Guide Strand Position 2′-15 Disulfide Peptide Conjugates Examples 99-100

Scheme 25 is shown in FIG. 25A to FIG. 25B-2.

Synthesis of N3-Seq 283 (Ex. 99)

RNA compound N2 (11 mg, 1.54 μmol; prepared as detailed in Section M for the di-click substrate) was dissolved in trifluoroethanol containing 50 mM acetic acid (1300 μL). To this solution was added peptide seq283 (3.57 mg, 2.31 μmol) dissolved in trifluoroethanol containing 50 mM acetic acid (1300 μL). The mixture was aged for 10 min, and purified by ion pairing chromatography on a Waters Xbridge phenyl column (10 μM, 19×250 mm) using a gradient of 5-80% acetonitrile/water with 100 mM triethylammonium acetate over 15 min at 20 mL/min. The product was isolated by spin dialysis against water (3×) followed by lyophilization to give a solid. MS (m/z): 8600.

Synthesis of N4-Seq 283-k (Ex. 100)

A solution of B2 (5.65 mg, 0.609 μmol) dissolved in DI water (423 μL) was added to N3-Seq 283 (5.24 mg, 0.609 μmol) and heated at 90° C. for 1 min. Solution was lyophilized to give duplex as a whilte solid. MS (m/z) passenger strand: 9268, guide strand:8601.

Additional Synthesis of N3 Peptide Conjugates and N4 Duplexes

Additional N3 conjugates of peptide sequences and the corresponding duplexes N4 were prepared in a manner analogous to that detailed above.

Section O. Synthesis of Guide Strand Position 2′-15 ECL Peptide Conjugates Examples 101-103

Scheme 26 is shown in FIG. 26A-1 to FIG. 26B-2.

Synthesis of O2 (Ex. 101)

RNA compound O1 (20.7 mg, 2.97 μmol; prepared in an analogous manner to L8) was dissolved in 100 mM NaHCO₃ (400 μL) and DMSO (300 μL). To this mixture was added a solution of 2,5-dioxopyrrolidin-1-yl 8-(((S)-3-methyl-1-oxo-1-(((S)-1-oxo-1-((4-((((2-(pyridin-2-yldisulfanyl)ethyl)carbamoyl)oxy)methyl)phenyl)amino)-5-ureidopentan-2-yl)amino)butan-2-yl)amino)-8-oxooctanoate L6 (6.28 mg, 7.43 μmol) dissolved in DMSO (250 μL). The reaction mixture was aged for 1.5 h, diluted with water (1.5 mL) and purified by ion pairing chromatography on a Waters Xbridge phenyl column (5 μM, 19×250 mm) using a gradient of 0-60% acetonitrile/water with 100 mM triethylammonium acetate over 15 min at 20 mL/min. The product was isolated by spin dialysis against water (3×) followed by lyophilization to give a solid. MS (m/z): 7696.

Synthesis of O2-Seq 463 (Ex. 102)

RNA compound O2 (10 mg, 1.30 μmol) was dissolved in trifluoroethanol containing 50 mM acetic acid (1000 μL). To this solution was added peptide Seq 463 (3.28 mg, 1.95 μmol) dissolved in trifluoroethanol containing 50 mM acetic acid (500 μL). The mixture was aged for 1 hr and purified by ion pairing chromatography on a Waters Xbridge phenyl column (5 μM, 19×250 mm) using a gradient of 5-90% acetonitrile/water with 100 mM triethylammonium acetate over 15 min at 20 mL/min. The product was isolated by spin dialysis against water (3×) followed by lyophilization to give a solid. MS (m/z): 9268.

Synthesis of O3-Seq 463-k (Ex. 103)

A solution of O2-Seq 463 (3.02 mg, 0.326 μmol) dissolved in DI water (303 μL) was added to B2 (3.02 mg, 0.326 μmol) and heated at 90° C. for 1 min. Solution was lyophilized to give duplex as a whilte solid. MS (m/z) passenger strand: 9267, guide strand:9264.

Additional Synthesis of O2 Peptide Conjugates and O3 Duplexes

Additional O2 conjugates of peptide sequences and the corresponding duplexes O3 were prepared in a manner analogous to that detailed above.

Section P. Synthesis of Guide Strand Position 2′-15 Cholesterol and Peptide Conjugates Examples 104-106

Scheme 27 is shown in FIG. 27A-1 to FIG. 27B-2.

Synthesis of P1 (Ex. 104)

RNA compound N2 (67.2 mg, 9.39 μmol) and diisopropylethylamine (13.1 μL, 75 μmol) was dissolved in water (750 μL), N,N-dimethylacetamide (750 μL), and tetrahydrofuran (1200 μL). To this mixture was added a solution of thiocholesterol (30.2 mg, 75 μmol) dissolved in tetrahydrofuran (300 μL). The mixture was aged for 30 min, diluted with 2M triethylammonium acetate (100 μL), and purified by ion pairing chromatography on a Waters Xbridge phenyl column (10 μM, 19×250 mm) using a gradient of 5-95% acetonitrile/water with 100 mM triethylammonium acetate over 15 min at 20 mL/min. The product was isolated by spin dialysis against water (3×) followed by lyophilization to give a solid. MS (m/z): 7451.

Synthesis of P2-Seq 32-k (Ex. 105)

A solution of P1 (1.0 mg, 0.134 μmol) dissolved in DI water (200 μL) was added to B10-Seq 32 (1.86 mg, 0.129 μmol) and heated at 90° C. for 1 min. Solution was lyophilized to give duplex as a whilte solid. MS (m/z) passenger strand: 13295, guide strand:7450.

Synthesis of P2-Seq 32-m (Ex. 106)

Guide strand P1 was also duplexed with passenger strand F6-Seq 32 in a manner identical to that detailed above in Example 105 to provide duplex P2-Seq 32-m:

Scheme 28 is shown in FIG. 28-1 to FIG. 28-2.

Section Q. 3′ Enzymatically Cleaved Linker Bis Peptides Examples 107-109

Scheme 29 is shown in FIG. 29A-1 to FIG. 29C-2.

Synthesis of Q1 (Ex. 107)

In a Falcon tube, L6 (13.82 mg, 0.016 mmol) was dissolved in DMSO (1963 μl) and cooled to 10° C. in an ice-bath. In a separate Falcon tube, B4 (76.2 mg, 8.18 μmol) was dissolved in pH 8.3 NaHCO₃ 200 mM (1309 μl). The RNA solution was added to the DMSO solution and the reaction was determined complete after 5 min.

The reaction was purified by ion-pairing chromatography (GX-281, XBridge Prep Phenyl 5 um, OBD, 30×150 mm, 30 mL/min, 5-45% of 100 mM TEAA in water/100 mM TEAA in ACN, 20 min gradient). The resulting fractions were dialyzed against water 3× on Millipore 3K, 15 mL tubes, (4200 rpm, 4° C.) and then lyophilized to afford a white solid. Expected mass: 10052.834. Found mass: 10051.0.

Synthesis of Q2-Seq 74 (Ex. 108)

See Synthesis of B10-Seq74 for reaction procedure. Q2-Seq 74-Found mass: 13940.012.

Synthesis of Q3-Seq 74-b (Ex. 109)

See Synthesis of B11-Seq74 for reaction procedure. Q3-Seq 74-b—Found mass: 20792.

Section R. 5′,3′ Di-Lipopeptide Conjugates Examples 110-112

Scheme 30 is shown in FIG. 30A to FIG. 30E-3.

Synthesis of R2 (Ex. 110)

L6 (23.2 mg) was dissolved in formamide (300 μl) and DMSO (300 μl), then added R¹ (50 mg) dissolved in pH 8.3 200 mM NaHCO₃ aqueous solution (600 μl). After 5 min, precipitation appeared. Additional DMSO (300 μl) was added, whereupon most of solids redissolved. After a 15 min incubation, the reaction was purified using an XBridge Prep Phenyl column (5 uM, 30×150 mm) using a gradient of 5-45% CH₃CN (100 mM TEAA)/water (100 mM TEAA), 20 min at 20 mL/min, collecting at 260 nm. The product fractions were diluted with water to reduce the CH₃CN content to below 20% and centrifugal dialyzed four times against water over a 3K membrane. The retentate was frozen and lyophilized to a white solid.

Synthesis of R3 (Ex. 111)

Dissolved R2 in 500 ul of water, dissolved Compound 35 of SCHEME 38 separately in 500 ul of water, then added GS solution to PS solution, vortexed thoroughly at RT, then checked analytical SAX HPLC confirming the formation of duplex. The solution was freeze dried to afford the duplex as a white amorphous solid.

Synthesis of R4-Seq 27-1 (Ex. 112)

Dissolved siRNA R3 in 2,2,2-trifluoroethanol containing 50 mM acetic acid (500 uL). Dissolved peptide in 2,2,2-trifluoroethanol containing 50 mM acetic acid (500 uL), then added 8 M aqueous guanidinium hydrochloride (30 uL). The siRNA solution was added to the peptide solution to give a clear solution. After 1 h, the reaction mix was diluted with formamide (1 mL) and was purified on neutral SAX system (Buffer A: 1:1 water:TFE 20 mM MES pH 5.5 Buffer B: 1:1 water:TFE 20 mM MES pH 5.5 1M CsCl) in two runs. The product fractions were diluted with water to reduce the TFE content to below 50% and dialyzed three times against water over a 3K membrane. The retentate was frozen and lyophilized to a white solid.

Additional Synthesis of R3 Peptide Conjugates and R4 Duplexes

Additional R3 conjugates of peptide sequences and the corresponding R4 duplexes were prepared in a manner analogous to that detailed above.

Section S. Preparation of Alternative TetraGalNAc Ligands Examples 113-115 Synthesis of TetraGalNAc Ligand Compounds 17a, 17b and 17c

The following Scheme 31 was used to prepare TetraGalNAc Compounds 17a, 17b and 17c.

Synthesis of Compound 13

To a solution of 5-chloro-1-pentanol (3.0 g, 24.47 mmol) Compound 11 in DMF (20 mL) was added sodium azide (1.909 g, 29.4 mmol) Compound 12. After being stirred at 60° C. for overnight, the reaction mixture was concentrated in vacu. The residue was purified by silica gel chromatography (EtOAc/Hexane 1:3), to give product Compound 13 as clear liquid. ¹HNMR (500 MHz, CDCl₃) δ 3.62 (m, 2H), 3.25 (t, J=6.9 Hz, 2H), 1.63-1.53 (m, 4H), 1.45-1.40 (m, 2H).

Synthesis of Compound 15

Compound 13 (0.796 g, 6.16 mmol) and D-galactosamine pentaacetate (2.00 g, 5.14 mmol) Compound 14 were suspended in 20 mL DCM, followed by addition of trifluoromethanesulfonic acid (0.154 g, 1.027 mmol). The resulting mixture was brought to reflux for overnight. LC-MS indicated completed conversion of SM, the reaction mixture was diluted with EtOAc and washed with sodium bicarbonate and dried over sodium sulfate. Solvent was removed and the residue was purified by ISCO DCM/MeOH from 100/0 to 90/10 over 30 min to afford Compound 15 as a white solid. ¹H NMR (500 MHz, CDCl₃) δ: 1.97 (6H, s), 2.02 (6H, s), 2.06 (6H, s), 2.15 (6H, s), 3.28 (6H, t, J=6.89 Hz), 3.50 (3H, dt, J=9.63, 6.66 Hz), 3.68 (1H, q, J=5.98 Hz), 3.94-3.92 (7H, m), 4.16-4.15 (5H, m), 4.73 (2H, d, J=8.34 Hz), 5.31 (2H, dd, J=11.16, 3.48 Hz), 5.40-5.38 (5H, m). Calculated mass: [M+H]+: C₁₉H₃₁N₄O₉, 459.2; observed: 459.4.

Synthesis of Compound 16

Lys-alkyne Compound A1 (130 mg, 0.436 mmol) and GalNAc Azide 6 (999 mg, 2.178 mmol) were dissolved in THF (5 mL, degassed). Copper (I) bromide-dimethyl sulfide complex (17.91 mg, 0.087 mmol) was added in one portion to the reaction mixture and the THF solution was stirred for overnight at 40° C. The reaction color changed to blue/green, indicating Cu²⁺, fresh sodium ascorbate 37 mg in 0.2 mL of water was added to reaction mixture and allowed to react overnight. The reaction was concentrated and purified by RP HPLC 5-60 MeCN (0.5% TFA)/Water (0.5% TFA) over 20 min. The collected fractions were combined and lyophilized to afford Compound 8 as a white solid. Calculated mass: [M+3H]³⁺: C₉₄H₁₄₅N₁₈O₃₈, 2134.0, m/z=711.3; observed: 711.9.

Synthesis of Compound 17a (Ex. 113)

To protected TetraGalNAc Compound 8 (300 mg, 0.141 mmol) in DCM/MeOH=1/1 5 mL at 0° C. was added Sodium Methoxide (91 mg, 1.688 mmol). The reaction was stirred for 1 h and quenched by addition of 2 mL of water. Volatile solvent was removed, and the reaction mixture was purified by P4 bio gel with water and the collect fractions were combined and lyophilized to afford Compound 9 as a white solid. Calculated mass: [M+3H]³⁺: C₇₀H₁₂₁N₁₈O₂₆, 1629.9, m/z=543.3; observed: 543.8; [M+2H]²⁺: C₇₀H₁₂₀N₁₈O₂₆, 1628.9, m/z=814.5; observed: 814.9.

Synthesis of Compounds 17b and 17c (Ex. 114 and Ex. 115)

Syntheses of Compounds 17b and 17c which have the following structures were accomplished in a manner similar to that used for Compound 17a using the appropriate azide source.

Scheme of Conjugation of TetraGalNAc Ligands

Scheme 32 as shown in FIG. 31A and FIG. 31B shows a general scheme that can be used to prepare tetraGalNAc-siRNA conjugates.

Using the general scheme 32, Conjugates 10-1, 10-2, 10-3, 10a-1, 17a-1, 17b-1, 17c-1 can be obtained. The coupling procedure can be performed on a preformed siRNA duplex or on a single strand followed by annealing. Alternatively, one can utilize the protocol outlined in Bioconjug Chem. 2011, 22, pp. 1723-8.

Example 117 Synthesis of TetraGalNAc-siRNA Conjugate (A11-a) via TetraGalNAc Acetate Compound A9

To a solution of tetraGalNAc acetate (A9, 58.7 mg, 0.027 mmol) in acetonitrile (1.5 ml) was added DIPEA (2.2 mg, 0.055 mmol) and HATU (10.44 mg, 0.027 mmol). The mixture was stirred at room temperature for 30 min, transferred into a solution siRNA (0.014 mmol) in water (1.5 ml) and acetonitrile (1.5 ml) via a syringe pump over 20 min, and stirred for 30 min before it was concentrated under vacuum down to 1.5 mL. Sodium carbonate (218 mg, 2.059 mmol) was then added, followed by MeOH (0.50 ml). The resulted solution was stirred at room temperature for 16 h, concentrated, purified via dialysis, and lyophilized to yield Conjugate A11-a.

The coupling protocol described for A11-a can also be performed with A10 instead of A9.

Examples 118-119 Synthesis of Conjugates A11-b and A11-c (Ex. 118 and Ex. 119)

A similar protocol was used for Conjugates A11-b and A11-c. Duplex formation with the appropriate antisense or sense strand can be performed using the protocol described for B11.

Example 120 Synthesis of 3′5′ Bis TetraGalNAc-siRNA Conjugate Single Strand 18

To a solution of tetraGalNAc acid Compound 10 (41.2 mg, 0.025 mmol) in DMSO (200 uL) was added HATU (9.6 mg, 0.025 mmol) and DIPEA (17.6 uL, 0.126 mmol). The mixture was stirred at room temperature for 15 min, transferred into a solution of diamino-siRNA (18.8 mg, 2.52 umol) in water (40 uL) and DMSO (360 uL) and stirred for 30 min. The mixture was diluted with water (1.5 mL) and purified on a XBridge Prep Phenyl column (5 uM, 19×250 mm) using a gradient of 0-30% CH₃CN/water containing 100 mM TEAA. The fractions were concentrated via dialysis and lyophilized to yield Compound 18.

Example 121 Synthesis of 3′5′ Bis TetraGalNAc-siRNA Duplex Conjugate 19-1 (Ex. 121)

Scheme 33 as shown in FIG. 32A and FIG. 32B was used to prepare TetraGalNAc-siRNA Conjugate 19-1.

A solution of 3′5′ bis tetraGalNAc-siRNA conjugate 18 (13.7 mg, 1.29 umol) in water (200 uL) was added to a solution of Guide siRNA (9.3 mg, 1.35 umol) dissolved in water (100 uL) and heated at 90 C for 1 minute. The resulting solution was cooled and lyophilized to yield duplex 19-1.

Example 122 Synthesis of TetraGalNAc Ligand Compound 24 (Ex. 122)

The following Scheme 34 was used to prepare tetraGalNAc ligand Compound 24.

Synthesis of Compound 22

To a solution of N—BOC-1,3-DIAMINOPROPANE (Compound 20, 115 mg, 0.660 mmol) in 1:1 CH₂Cl₂/CH₃CN (1 mL) at 0° C. was added a solution of 3-maleimidopropionic acid N-hydroxysuccinimide ester (Compound 21, 185 mg, 0.695 mmol) dissolved in acetonitrile (4 mL) and CH₂Cl₂ (1 mL). The mixture was stirred for 1 h and concentrated in vacuo. The residue was purified by silica gel chromatography (0-5% MeOH/CH₂Cl₂ to give product Compound 22. Calculated mass: [M+H]+: C₁₅H₂₄N₃O₅, 326.2; observed: 326.3.

Synthesis of Compound 23

To a solution of maleimide Compound 22 (56 mg, 0.172 mmol) in CH₂Cl₂ (1 ml) was added a solution of 4M HCl (1 ml, 4.00 mmol) in dioxane. The mixture was stirred for 1 h and concentrated in vacuo. The residue was azeotroped with CH₂Cl₂ (2×) and dried under vacuum to give product Compound 23. Calculated mass: [M+H]+: C₁₀H₁₆N₃O₃, 226.1; observed: 226.3.

Synthesis of tetraGalNAc maleimide Compound 24 (Ex. 122)

To a solution of tetraGalNAc acid Compound 10 (100 mg, 0.061 mmol) in DMF (500 uL) was added HATU (34.9 mg, 0.092 mmol), Et₃N (42.6 uL, 0.306 mmol) and N-(3-aminopropyl)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanamide hydrochloride (16.0 mg, 0.061 mmol). The mixture was stirred at room temperature for 1.5 h, acidified with TFA and purified by reverse phase 0-50% CH₃CN/water containing 0.1% TFA. The fractions were lyophilized to yield Compound 24. Calculated mass: [M+2H]²⁺: C₇₆H₁₂₅N₂₁O₃₂, 1843.8, m/z=921.9; observed: 922.7.

Example 123 Synthesis of Compound 26

Scheme 35 as shown in FIG. 33A and FIG. 33B was used to prepare Compound 26.

To a degassed solution of 2′-3,17 propargyl siRNA (RNA 25, 33 mg, 4.49 umol) and PEG9 SPDP azide (26 mg, 36 umol, prepared from commercial PEG-azide and pyridyl disulfide reagents) in 3:1 DMA/water (1 mL) was added a degassed solution of Copper (I) Bromide-Dimethylsulfide Complex (1.8 mg, 9.0 umol). The mixture was stirred for 72 h at room temperature, diluted with water (2 mL), filtered using a 0.45 uM syringe filter and concentrated by dialysis. The concentrated mixture was purified on a XBridge Prep Phenyl column (5 uM, 19×250 mm) using a gradient of 0-50% CH₃CN/water containing 100 mM TEAA. The fractions were concentrated via dialysis and lyophilized to yield Compound 26.

Examples 124-125 Synthesis of Compounds 27 and 28 (Exs. 124-125)

Scheme 36 as shown in FIG. 34A to FIG. 34C was used to prepare Compounds 27 and 28.

Synthesis of Compound 27 (Ex. 124)

To a solution of 2′-3,17 click PEG9 SPDP Conjugate 26 (13.2 mg, 1.50 μmol) in water (1 mL) was added a solution of TCEP hydrochloride (9.15 mg, 32.2 umol) dissolved in water (0.5 mL). The mixture was stirred at RT for 30 min then purified on a XBridge Prep Phenyl column (5 uM, 19×250 mm) using a gradient of 5-40% CH₃CN/water containing 100 mM TEAA. The fractions were concentrated via dialysis and lyophilized to yield Compound 27.

Synthesis of Compound 28 (Ex. 125)

To a solution of 2′-3,17-click PEG9SH 27 (3 mg, 0.35 μmol) in pH 6.0 acetate buffer (100 uL) was added a solution of tetra GalNAc maleimide (5.1 mg, 2.77 μmol) dissolved in pH 6.0 acetate buffer (100 uL). The mixture was stirred at room temperature for 30 min then purified on a XBridge Prep Phenyl column (5 uM, 19×250 mm) using a gradient of 5-40% CH₃CN/water containing 100 mM TEAA. The fractions were concentrated via dialysis and lyophilized to yield Compound 28.

Example 126 Synthesis of 2′-3,17 Bis TetraGalNAc-siRNA Duplex Conjugate 29

The procedure detailed for Conjugate 19 was used to duplex 28 to make Conjugate 29.

Example 127 Synthesis of TetraGalNAc Thiol Compound 31

Scheme 37 below was used to prepare Compound 31.

To a solution of tetraGalNAc acid Compound 10 (54 mg, 0.033 mmol) in N,N-dimethylacetamide (500 μl), was added cystamine dihydrochloride 30 (14.9 mg, 0.066 mmol), EDC (12.7 mg, 0.066 mmol), HOAT (10.2 mg, 0.066 mmol) and DIPEA (57.7 μl, 0.330 mmol). The mixture was stirred at room temperature for 18 h, then added a solution of DTT (50.9 mg, 0.330 mmol) in N,N-dimethylacetamide (100 μl). The mixture was stirred at room temperature for 0.5 h, acidified with TFA and purified by reverse phase 0-30% CH₃CN/water containing 0.1% TFA. The fractions were lyophilized to yield Compound 31. Calculated mass: [M+2H]²⁺: C₆₈H₁₁₅N₁₉O₂₉S, 1695.8, m/z=847.9; observed: 848.0.

Examples 128-130 Synthesis of Conjugates 35-37

Scheme 38 as shown in FIG. 35A and FIG. 35B was used to prepare Conjugates 35-37.

Synthesis of Compound 33

To a degassed solution of 2′-click 15 GS Compound 32 (130 mg, 0.019 mmol) and (9H-fluoren-9-yl)methyl (2-azidoethyl)carbamate (29.1 mg, 0.095 mmol) in 3:1 DMA/water (2 mL) was added a solution of Copper (I) bromide-dimethylsulfide Complex (9.72 mg, 0.042 mmol) dissolved in degassed DMSO (0.32 mL). The mixture was stirred at 45° C. for 2 h, cooled to room temperature, and added pH 8 EDTA (0.5 M, 2 mL) to quench reaction. Stirred for 15 min and purified on a XBridge Prep Phenyl column (5 uM, 30×150 mm) using a gradient of 0-45% CH₃CN/water containing 100 mM TEAA. The fractions were concentrated via dialysis. To the combined material in water (3 mL) was added a solution of piperidine (936 μL, 1.891 mmol). The mixture was stored at 4° C. for 18 h, diluted with water (10 mL) and filtered off solids through syringe filter. Added pH 8 EDTA (0.5 M, 2 mL), concentrated via dialysis and lyophilized to yield Compound 33.

Synthesis of Compound 34

To a solution of 2′-15 click C2 NH2 GS Compound 33 (43.6 mg, 6.26 μmol) in 200 mM NaHCO3 soln (2000 μl) and formamide (1000 uL) was added a solution of N-Succinimidyl-3-[2-pyridyldithio]propionate (17.9 mg, 0.057 mmol) dissolved in DMSO (298 uL). The mixture was stirred at 10° C. for 15 min, diluted with water (10 mL) and Formamide (1 mL), and concentrated by dialysis. Added 2M TEAA (200 uL) and purified on a XBridge Prep Phenyl column (5 uM, 19×250 mm) using a gradient of 5-40% CH₃CN/water containing 100 mM TEAA. The fractions were concentrated via dialysis and lyophilized to yield Compound 34.

Synthesis of 2′-15 TetraGalNAc-siRNA Conjugate 35 (Ex. 128)

To a solution of 2′-15 click C2 NH2 NHS SPDP GS Compound 34 (13 mg, 1.82 μmol) in 1:1 formamide/water (200 μl) was added a solution of tetraGalNAc SH (4.62 mg, 2.72 mol) in formamide (200 uL). The mixture was stirred at room temperature for 3.5 h, added 2M TEAA (50 uL) and purified on a XBridge Prep Phenyl column (5 uM, 19×250 mm) using a gradient of 2-35% CH₃CN/water containing 100 mM TEAA. The fractions were concentrated via dialysis and lyophilized. The resulting solid was purified on a Proteomix SAX-NP10 column (22.1×50 mm) using a gradient of 2-30% (Solvent A: 60:40 TFE/water with 40 mM Et3N, Solvent B: 60:40 TFE/water with 40 mM Et3N, 1M Guanidine HCl). The fractions were concentrated via dialysis and lyophilized to yield Conjugate 35.

Synthesis of Conjugates 36 and 37 (Ex. 129 and Ex. 130)

The procedure detailed for Conjugate 19-1 was used to duplex Conjugate 35 and the appropriate passenger strand to prepare Conjugates 36 and 37, respectively.

Examples 131-139 Synthesis of Conjugates 38-45 (Exs. 131-139)

Scheme 39 as shown in FIG. 36A to FIG. 36C, was used to prepare Conjugates 38-44.

Scheme 40. Examples of different linkers from Table 2 as shown in FIG. 37, used to conjugate tetraGalNAc to siRNA.

Step 1: Passenger-RNA and Linker, Example with Proline to Illustrate Protocol

To a solution of FMOC—PRO-OH (11.11 mg, 0.033 μmol) in 120 μL DMSO were added DIPEA (43.2 μl, 0.247 μmol) followed by HATU (10.96 mg, 0.029 μmol). The mixture, slightly yellow, was stirred at room temperature for 30 min. The mixture was then added to a solution of the oligonucleotide passenger strand TEAA salt (60 mg, 8.24 μmol) in 500 μL of (10% H₂O/DMSO), and the mixture continued to stir at room temperature for one hour. The reaction mixture showed desired product via LC-MS. To the reaction mixture was added diethylamine (43.0 μl, 0.412 μmol) and the mixture was stirred for one hour, confirmed desired product via LC-MS. The reaction mixture was purified by centrifugal dialysis using 3 kDa cut-off membrane. The process was repeated three times with water (14 mL each time). The resulting solution was concentrated, frozen, and lyophilized overnight to yield product as a white fluffy solid. LC/MS confirms product [7384.9].

Step 2: TetraGalNAc-Linker-Passenger RNA

To a solution of TetraGalNAc Compound 10 (53.2 mg, 0.033 μmol) in 532 μL DMSO were added DIPEA (42.6 μl, 0.244 μmol) followed by HATU (12.36 mg, 0.033 μmol). The mixture, slightly yellow, was stirred at RT for 30 min. The mixture was then added to a solution of the linker-oligonucleotide passenger strand in 500 μL of DMSO, and the mixture continued to stir at room temperature for two hours. LC/MS showed desired product. The reaction mixture was subjected to centrifugal dialysis using 3 kDa cut-off membrane. The process was repeated three times with water (14 mL each time). The resulting solution was purified by Gilson PLC 2020 using XBRIDGE PHENYL, 10-27% CH₃CN with 200 μM TEAA for 35 minutes. Collection solution was concentrated via centrifugal dialysis using 3 kDa cut-off membrane. The resulting concentrated solution was treated with 1.0N NaCl and centrifugal dialysis. The process was repeated five times with water (14 mL each time). The resulting concentrated solution (˜1.5 mL) was frozen and lyophilized overnight to yield product as a white fluffy solid. LC/MS confirms product [9002.5].

Step 3: Duplex Formation

To a TetraGalNAc-linker-RNA (18.5 mg, 2.055 μmol) in 1.5 mL of water was duplexed with ApoB guide strand (14.12 mg, 2.055 μmol) in 1.5 mL of water. The mixture was heated at 90° C. for 5 min with stir bar. The duplex was cooled and stir bar removed. The solution was lyophilized over two days to yield desired duplex Conjugate 38 as a white fluffy solid. LC/MS confirms product [16048].

ALL the remaining conjugates were prepared using the same general procedure.

Examples 140-142 Synthesis of Compounds/Conjugates 46-48

Scheme 41 as shown in FIG. 38A to FIG. 38E was used to prepare Compounds and/or Conjugates 46-48.

Synthesis of RNA Compound 46 (Ex. 140)

SPDP Acid (2.2 mg, 10.3 μmol) was dissolved DMSO 100 μL and N,N-diisopropylethylamine (14.0 μl, 0.08 mmol), HATU (19.6 mg, 0.051 mmol) were added sequentially. RNA (15 mg, 2.06 μmol) in 200 μL of DMSO:Water (9:1) was added and the resulting reaction mixture was stirred for 1 h, reaction was quenched by addition of 3 mL water and dialyzed down to 500 μL, diluted by formamide to 3 mL and purified by SAX (Buffer A: 60% TFE in water, 20 mM TEA, Buffer B: 60% TFE in water, 20 mM TEA, 1 M CsCl, gradient A/B from 100/0 to 35/65 over 15 min). The collected fractions were combined and dialyzed against water and lyophilized to afford Compound 46 as a white solid. Calculated mass: [M−H]⁻: C₂₃₄H₃₀₀F₈N₇₂O₁₅₀P₂₃S₃, 7480.1; observed: 7483.0.

Synthesis of Conjugate 47 (Ex. 141)

RNA Compound 46 (22 mg, 2.9 μmol) and tetraGalNAc Thiol Compound 31 (10.0 mg, 5.9 μmol) were dissolved in formamide:pH=6.8 Tris buffer (3:1) 400 μL and stirred for 1 h. The reaction mixture was purified by SAX (Buffer A: 60% TFE in water, 20 mM TEA, Buffer B: 60% TFE in water, 20 mM TEA, 1 M CsCl, gradient A/B from 100/0 to 35/65 over 15 min). The collected fractions were combined and dialyzed against water and lyophilized to afford Conjugate 47 as a white solid. Calculated mass: [M−H]⁻: C₂₉₇H₄₁₀F₈N₉₀O₁₇₉P₂₃S₃, 9063.9; observed: 9066.2.

Synthesis of Conjugate 48 (Ex. 142)

Conjugate 47 (10.9 mg, 1.20 μmol) and guide strand (7.81 mg, 1.14 μmol) were mixed in RNAse free water 1 mL for 2 h. The reaction mixture was lyophilized to afford duplex Conjugate 48 in quantitative yield.

Examples 143-145 Synthesis of Compounds/Conjugates 49-51

Scheme 42 as shown in FIG. 39A to FIG. 39C was used to prepare Compounds and/or Conjugates 49-51.

Synthesis of RNA Compound 49 (Ex. 143)

33.3 mg of siRNA passenger strand was weighed into a 4 mL vial then 1 mL 100 mM NaHCO3 was added to dissolve. Added 0.86 uL of propionic anhydride and let stir at RT. After aging ˜2 h, spin dialyzed 3× against water. Filtered through frit and the solution was dried via lypophilization to afford RNA Compound 49.

Synthesis of Conjugate 50 (Ex. 144)

Step 1. Charge 2.8 mg azide, 25.7 mg siRNA, 25 ml N2 sparged DMSO and 4 ml water to 40 mL vial. Sparge with N₂. Charge 2.98 mL of Cu/ligand solution (N₂ sparged, 20/100 umol in 10 ml DMSO). Agitate at RT under sparged N₂.

Step 2. Charge Compound 10 and 1 ml DMSO. Charge 6 uL of DIPEA and agitate for 2 min. Charge 6 mg HBTU and agitate for 2 min. Charge siRNA mixture from Step 1. The reaction was not complete so repeated with half of previous reagent charge. Evaporated the reaction mixture, dialyzed and HPLC purified (X-Bridge Phenyl, TEAA/ACN gradient). Evaporated, dialyze and lyophilized to afford Conjugate 50.

Synthesis of Conjugate 51 (Ex. 145)

Dissolve GS (Conjugate 50) 10.65 mg in 1 ml water and dissolve PS (Conjugate 49) 10.20 mg in 1.17 ml water. Added 8.7 mg of Conjugate 49 to all of Conjugate 50 to form a 1:1 duplex. Heat to 90° C. for 1 min, cool to RT over 15 min. The solution was filtered and dried via lyophilization to afford Conjugate 51 as a white solid.

RNA Silencing Activity of Compounds Transfected with Lipofectamine in Luciferase Constructs

HEK293 cells stably transfected with luciferase vector that contains target sites for siRNA in 3′UTR of renilla luciferase were generated. These cells were seeded on 96-well tissue culture plates (Corning: #3903) at a density of 7.5e3 cells per well in DMEM 10% serum media. Cellular plates were then incubated at 37° C./5% CO2 for 24 hr. After incubation, plates were treated with test compounds co-transfected with transfection reagent Lipofectamine 2000 (invitrogen: #11668-O19) in Opti-MEM (Gibco: #31985) in accordance to manufacturers protocol. The treatment concentrations ranged from 10 nM to 0.03 pM. Treated plates were then incubated for 24 hr at 37° C./5% CO2. Following treatment incubation, cells were lysed and processed in accordance to Dual-Glo™ Luciferase Assay (Promega: E2920) and read on a TECAN safire2 μlate reader.

RNA Silencing Activity of Compounds Transfected with Lipofectamine in HepG2 Cells

HepG2 cells (ATCC: HB-8065) were seeded on collagen coated plates (BioCoat: 356649) at a density of 7.5e3 cells per well in DMEM 10% serum media. Cellular plates were then incubated at 37° C./5% CO2 for 24 hr. After incubation, plates were treated with test compounds co-transfected with transfection reagent Lipofectamine 2000 (invitrogen: 11668-019) in Opti-MEM (Gibco: 31985) in accordance to invitrogen protocol. The treatment concentrations ranged from 10 nM to 0.03 pM. Treated plates were then incubated for 24 hr at 37° C./5% CO2. Following treatment incubation, cells were lysed with PLA Buffer (AB: 4448542) in accordance to supplied protocol. Resulting cell lysate was reverse transcribed to cDNA using High Capacity cDNA Kit (AB: 4368813) and run through qPCR using Life Technology 7900.

In Vivo Evaluation of RNAi Activity

CD1 female mice were dosed by subcutaneous injection in 200 ul volume. Animals were observed for behavioral or physiological changes. Animals were sacrificed 72 hrs post dose by CO2 asphyxiation followed by ex-sanguination via cardiac puncture. The liver samples were as 3 mm punches from the medial lobe and put into RNAlater tubes for isolation of total RNA. The mRNA knockdown analysis was conducted by Taqman analysis using standard procedures.

Scheme 43. General Description for Illustrative Purposes of Nomenclature Used in Table 6 as shown in FIG. 40. Exact siRNA sequences used in Table 6 can be found in Table 5.

A summary of in vitro and in vivo data of selected Compounds/Conjugates is shown in Table 6 and Table 7.

TABLE 6 In vitro and In Vivo Activity for Compounds Described in Section B-D. RBC Hemolysis Data on Free Peptide % KD % KD % KD EC 50 EC 50 2.5 mpk 5 mpk 2.5 mpk pH7.4 pH5.5 (SC (iv (iv Compound # (uM) (uM) admin) admin) admin) B8-seq137-b 8.3 4.3 47 B8-seq470-b 8.5 3.8 57 B8-seq1678-b >20 5 49 B8-seq 92-b 0.3 0.3 57 B8-seq1677-b 10 0.4 57 B8-seq-463-b 18 9.8 61 B8-seq1675-b 7 4.5 47 B11-seq1-b 5.3 0.7 49 B11-seq2-b >10 1.2 32 B11-seq3-b >10 0.5 49 B11-seq4-b 4.3 0.2 55 B11-seq5-b 5 0.5 74 B11-seq6-b >10 1 53 B11-seq7-b >10 0.7 45 B11-seq8-b 22 B11-seq9-b 8.9 1.7 28 B11-seq10-b 6 1.8 35 B11-seq11-b 0.39 0.04 21 B11-seq12-b 2 0.2 45 B11-seq13-b 1.9 0.2 5 64 B11-seq14-b 2.27 1.61 26 B11-seq15-b >10 0.4 28 B11-seq16-b 2.8 0.6 26 B11-seq17-b 4.4 0.7 34 B11-seq18-b 1 0.4 61 B11-seq19-b >10 0.7 64 B11-seq20-b 3.7 2.05 63 B11-seq21-b 2.2 0.4 56 B11-seq22-b 6 0.5 33 B11-seq23-b 7.3 6.1 59 B11-seq24-b >10 0.2 58 B11-seq25-b >10 3.6 52 B11-seq26-b 4.6 1.4 38 65 57 B11-seq27-b >10 0.4 61 B11-seq28-b 0.7 0.1 25 B11-seq29-b >10 2 20 B11-seq30-b >10 1.5 29 B11-seq31-b 1.5 0.3 64 B11-seq32-b 4.5 1.4 58 B11-seq33-b 0.02 0.04 35 B11-seq34-b 0.12 0.05 30 B11-seq35-b 0.03 0.03 37 B11-seq36-b 7.5 2.5 53 B11-seq37-b 6 2 22 B11-seq38-b 0.95 0.44 61 B11-seq39-b 1 0.6 58 B11-seq40-b 0.2 0.2 63 B11-seq41-b >10 0.7 36 27 B11-seq42-b 1.3 1.9 41 57 B11-seq43-b 0.9 0.3 55 B11-seq44-b 2.1 1.4 33 56 B11-seq45-b >10 0.07 51 53 B11-seq46-b 1.1 0.04 56 46 B11-seq47-b >10 0.4 49 51 B11-seq48-b 3.1 1.5 47 61 B11-seq49-b 4 0.6 37 49 B11-seq50-b >10 1.9 10 43 B11-seq51-b 11 48 B11-seq52-b >10 6.4 14 59 B11-seq53-b 1.17 0.37 45 B11-seq54-b 0.89 0.38 49 B11-seq55-b 0.51 0.18 −7 47 B11-seq56-b 1.46 0.19 12 48 B11-seq57-b 3.5 0.59 −11 B11-seq58-b 14.47 0.31 18 B11-seq59-b >20 0.65 7 52 B11-seq60-b 19.57 0.38 39 B11-seq61-b 1.39 0.65 55 B11-seq62-b >20 5.86 52 B11-seq63-b 0.94 0.64 37 B11-seq64-b >20 1.8 41 B11-seq65-b 1.38 1.87 28 B11-seq66-b >20 0.82 54 B11-seq67-b >20 0.87 39 B11-seq68-b >20 5.05 56 B11-seq69-b >20 0.91 34 B11-seq70-b 3.68 1.86 32 B11-seq71-b >20 3.56 44 B11-seq72-b 10.63 2.54 39 B11-seq73-b >20 4.2 38 B11-seq74-b 12.68 4.34 60 B11-seq75-b >10 0.9 55 B11-seq76-b 6.4 1.7 3 53 B11-seq77-b 0.17 0.23 38 B11-seq78-b 0.2 0.33 47 B11-seq79-b 1.52 1.86 47 B11-seq80-b >20 6.24 56 B11-seq81-b >20 3.91 51 B11-seq82-b 17 1.79 40 B11-seq83-b >20 6.19 35 B11-seq84-b 0.7 0.15 44 B11-seq85-b >10 0.1 45 B11-seq86-b >20 17.81 27 B11-seq87-b >10 0.02 30 B11-seq88-b 2.35 0.07 56 B11-seq89-b 3.29 0.14 51 B11-seq90-b >10 0.5 42 B11-seq91-b 26 B11-seq92-b 59 B11-seq93-b >20 5.88 51 B11-seq94-b 5.2 1.61 46 B11-seq95-b 3.59 3.1 43 B11-seq96-b 16.08 4.9 55 B11-seq97-b >20 5.56 52 B11-seq98-b >20 3.37 40 B11-seq99-b 12.9 5.61 43 B11-seq100-b 10.24 3.45 43 B11-seq101-b >20 4.85 46 B11-seq102-b >20 4.87 54 B11-seq103-b >20 3.86 43 B11-seq104-b 6.72 3.26 56 B11-seq105-b >10 >10 30 B11-seq106-b 8.4 0.24 34 B11-seq107-b 10.41 3.52 41 B11-seq108-b 5.6 2.69 40 B11-seq109-b >20 5.78 36 B11-seq110-b >20 3.36 43 B11-seq111-b >20 0.26 36 B11-seq371-b >20 2.8 45 B11-seq-1675-b 14.2 3.5 53 B13-seq 1676-b 14.2 3.5 53 B8-seq32-c 4.5 1.4 C6-seq-31c 1.5 0.3 31 C6-seq32-c 4.5 1.4 36 C6-seq106-c 7 0.7 30 C12-seq32-c 4.5 1.4 68 C15-seq32-c 4.5 1.4 39 D7-seq32-d 4.5 1.4 52 E10-seq 137-b >20 3.3 F6-seq 26-f >20 >20 47 F6-seq32-f 4.5 1.4 47 F6-seq463-f 18 9.8 60 F6-seq491-f >20 3.3 72 F6-seq492-f >20 6.3 66 F6-seq-612-f 19 6 59 F6-seq1693-f 17.1 0.6 38 F6-seq1694-f 15.6 4.4 43 G5-seq463-g 18 9.8 47 G5-seq489-g >20 >20 48 H7-seq8-h 20 1.3 13 25 H7-seq26-h 4.6 1.4 35 H7-seq32-h 4.5 1.4 20 30 H7-seq37-h 6 2 39 H10-seq26-h 4.6 1.4 20 H10-seq32-h 4.5 1.4 33 I10-seq-1680-f >20 1.6 67 I10-seq-1681-f >20 1.4 66 I10-seq-1682-f >20 1.6 66 K6-seq37-h 6 2 55 K6-seq-74-h 12.7 4.3 48 K6-seq463-h 18 9.8 55 L11-seq 463j 18 9.8 52 M4-seq463-j 18 9.8 52 N4-seq106-k 7 0.7 69 N4-seq197-k >20 >20 63 N4-seq283-k >20 >20 64 O3-seq-463-k 18 9.8 35 70 P2-seq32-k 4.5 1.4 61 P2-seq32-m 4.5 1.4 64 Q3-seq 32-b 4.5 1.4 45 Q3-seq 74-b 12.7 4.3 43 Q3-seq 1675-b 14.2 3.5 70 R4-seq1690-l 1.9 0.6 79 R4-seq1691-l 1.6 0.5 55 R4-seq1692-l >20 >20 72 R4-seq1695-l 14.2 0.3 79 R4-seq1696-l >20 >20 36

TABLE 7 In vitro and In Vivo Activity for Compounds Generated in Section E. (Starting siRNA sequence information can be found in Table 8). Starting IC50 w/ siRNA Dose (mpk) LF2K ASGR sequence Route of In vivo % KD in HEK- binding Entry Compound code Administration (72 h) Luc [pM] IC50 nM 1 10a-1 51 5, 15 SC 33.6; 69.5 15.44 36.7 2 10b-1 54 SC 5, 15; IV 15 42, 49, 13 19.64 18.1 3 10-2 56 5, 50 SC 40, 56 (24 h) 23.4 4 10-3 57 1, 2.5, 5 SC 20, 45, 60 52 (HepG2) 5 17a-1 51 5 SC; 15 IV 11, 5 20.16 49.1 6 17b-1 54 5 SC; 15 IV 12, 22 43.96 33.3 7 19-1 52 5; 15 SC 32; 68 24.04 3.6 8 29 53 15 SC; 15 IV 43, 0 17.83 22 9 36 58 1, 2.5, 5 SC 16, 43, 56 10 37 58 1, 2.5, 5 SC 16, 32, 40 11 38 51 5 SC, 15 IV 36, 33 71 17 12 39 51 5 SC, 15 IV 19, 31 46.8 44 13 40 51 5, 15 SC 33, 62 76.8 77 14 41 51 5, 15 SC 28, 74 98.6 134 15 42 51 5, 15 SC 19, 73 309.7 135 16 43 51 5, 15 SC 8, 73 64.8 45 17 44 51 5, 15 SC 31, 73 67.1 66 18 45 51 5 SC, 15 IV 20, 4 73.4 11 19 48a-1 51 5, 15 SC 10.24; 59.93 23.43 20 48b-1 53 5, 15 SC 19.87; 42.08 57.96 21 51 55 5; 15 40; 45 1838.47 94.8

TABLE 8 Starting siRNA sequence information used to prepare conjugates from Table 7. Gene Duplex SEQ ID Entry Target Strand Sequence Code NO.: 1 ApoB Passenger [6amiL][iB][omeC][omeU][omeU][omeU][fluA][fluA] 51 1721 [omeC][fluA][fluA][omeU][omeU][omeC][omeC][omeU] [fluG][fluA][fluA][fluA][omeU][dTs]dT[iB] ApoB Guide [rAs][rUs][rUs][omeU][omeC][fluA][fluG][fluG][fluA] 1722 [fluA][omeU][omeU][fluG][fluU][omeU][fluA][fluA][fluA] [fluG][omeUs][omeU] 2 ApoB Passenger [6amiL][iB][omeC][omeU][omeU][omeU][fluA][fluA] 52 1723 [omeC][fluA][fluA][omeU][omeU][omeC][omeC][omeU] [fluG][fluA][fluA][fluA][omeU][dTs]dT[iB][6amiL] ApoB Guide [rAs][rUs][rUs][omeU][omeC][fluA][fluG][fluG][fluA] 1724 [fluA][omeU][omeU][fluG][fluU][omeU][fluA][fluA][fluA] [fluG][omeUs][omeU] 3 ApoB Passenger [6amiL][iB][omeC][omeU][clickU][omeU][fluA][fluA] 53 1725 [omeC][fluA][fluA][omeU][omeU][omeC][omeC][omeU] [fluG][fluA][clickA][fluA][omeU][dTs]dT[iB][C6SH] ApoB Guide [rAs][rUs][rUs][omeU][omeC][fluA][fluG][fluG][fluA] 1726 [fluA][omeU][omeU][fluG][fluU][omeU][fluA][fluA][fluA] [fluG][omeUs][omeU] 4 ApoB Passenger [iB][omeC][omeU][omeU][omeU][fluA][fluA][omeC] 54 1727 [fluA][fluA][omeU][omeU][omeC][omeC][omeU][fluG] [fluA][fluA][fluA][omeU][dTs]dT[iB][6amiL] ApoB Guide [rAs][rUs][rUs][omeU][omeC][fluA][fluG][fluG][fluA] 1728 [fluA][omeU][omeU][fluG][fluU][omeU][fluA][fluA][fluA] [fluG][omeUs][omeU] 5 ApoB Passenger [6amiL][iB][omeC][omeU][omeU][omeU][fluA][fluA] 55 1729 [omeC][fluA][fluA][omeU][omeU][omeC][omeC][omeU] [fluG][fluA][fluA][fluA][omeU][dTs]dT[iB] ApoB Guide [rAs][rUs][rUs][omeU][omeC][fluA][fluG][fluG][fluA] 1730 [fluA][omeU][omeU][fluG][fluU][clickU][fluA][fluA][fluA] [fluG][omeUs][omeU] 6 SSB Passenger [6amiL][iB][fluA][omeC][fluA][fluA][omeC][fluA][fluG] 56 1731 [fluA][omeC][omeU][omeU][omeU][fluA][fluA][omeU] [fluG][omeU][fluA][fluA][dTs]dT[iB] SSB Guide [rUs][rUs][rAs][omeC][fluA][omeU][omeU][fluA][fluA] 1732 [fluA][fluG][omeU][omeC][fluU][fluG][omeU][omeU][fluG] [omeU][omeUs][omeU] 7 CTNNB1 Passenger [6amiL][iB][omeC][omeU][clickG][omeU][omeU][fluG] 57 1733 [fluG][fluA][omeU][omeU][fluG][fluA][omeU][omeU] [omeC][fluG][clickA][fluA][fluA][omeUs][omeU][iB][C3SH] CTNNB1 Guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1734 [omeC][fluA][omeA][fluU][omeC][fluC][omeA][fluA] [omeC][fluA][omeG][omeUs][omeU] 8 CTNNB1 Passenger [6amiL][iB][omeC][omeU][clickG][omeU][omeU][fluG] 58 1735 [fluG][fluA][omeU][omeU][fluG][fluA][omeU][omeU] [omeC][fluG][clickA][fluA][fluA][omeUs][omeU][iB][C3SH] CTNNB1 Guide [omeUs][fluUs][omeUs][fluC][omeG][fluA][omeA][fluU] 1736 [omeC][fluA][omeA][fluU][omeC][fluC][clickA][fluA] [omeC][fluA][omeG][omeUs][omeU] As used herein, ome = 2′ methoxy; flu = 2′ fluoro; click = 2′ propagyl; iB = inverted abasic; “s” subscript = phosphorothioate; and r = 2′ ribo; 6amil = n-hexylamino; C3SH = n-propylthiol; and C6SH = n-hexylthiol.

One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The methods and compositions described herein, as presently representative of preferred embodiments, are exemplary and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art, which are encompassed within the spirit of the invention, are defined by the scope of the claims. 

What is claimed is:
 1. A modular composition comprising: 1) a single stranded or double stranded oligonucleotide; 2) one or more tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different:

wherein X is —O—, —S—, —CR¹R²— or —NR¹—, wherein R¹ and R² are each independently selected from the group consisting of hydrogen and C1-C6 alkyl; n is 1, 2, 3, or 4; and the bond with “

” indicates the point of attachment; optionally, 3) one or more linkers, which may be the same or different; optionally, 4) one or more peptides independently selected from SEQ ID No. 1-474, or the D-amino acid, retro-inverso, and cysteine conjugation point variants thereof, which may be the same or different, wherein the cysteine conjugation point variants thereof refers to variants of the peptides comprising conjugation through existing cysteines or through a cysteine residue added to a N— or C— terminus of the peptides; and optionally, 5) one or more targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.
 2. The modular composition of claim 1 comprising: 1) a single stranded or double stranded oligonucleotide; 2) 1-8 tetraGalNAc ligands of Formula (II), which may be the same or different, wherein X is —O—, —S—, —CH₂— or —NH—; and n is 1, 2, 3, or 4; 3) 1-24 linkers, which may be the same or different; 4) 1-8 peptides independently selected from SEQ ID No. 1-474, which may be the same or different; and optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.
 3. A modular composition comprising: 1) a single stranded or double stranded siRNA; 2) 1-8 tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different, wherein X is —O—, —S—, —CH₂— or —NH—; and n is 1, 2, 3, or 4; 3) 1-24 linkers, which may be the same or different; 4) 1-12 peptides independently selected from SEQ ID No. 1-474, or the D-amino acid, retro-inverso, and cysteine conjugation point variants thereof, which may be the same or different, wherein the cysteine conjugation point variants thereof refers to variants of the peptides comprising conjugation through existing cysteines or through a cysteine residue added to a N- or C-terminus of the peptides; and optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.
 4. The modular composition of claim 3, wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA optionally via linkers.
 5. The modular composition of claim 3, wherein X of Formula (I), (II) or (III) is —O—, —S— or —CH₂—; and n is 1, 2 or
 3. 6. The modular composition of claim 3, wherein the composition comprises 1-4 tetraGalNAc ligands, which may be the same or different.
 7. The modular composition of claim 3, wherein the composition comprises 1-8 peptides, which may be the same or different.
 8. The modular composition of claim 3, wherein the siRNA is double stranded; and wherein the tetraGalNAc ligands are attached to the guide strand or the passenger strand of the siRNA at different 2′-positions of the ribose rings of the siRNA.
 9. The modular composition of claim 3, wherein the siRNA is double stranded; and wherein the tetraGalNAc ligands are attached to the guide strand or the passenger strand of the siRNA at different terminal 3′ and/or 5′-positions.
 10. The modular composition of claim 3, wherein the siRNA is double stranded; and wherein the tetraGalNAc ligands are attached to both the guide strand and the passenger strand of the siRNA at different 2′-positions of the ribose rings and/or different terminal 3′ and/or 5′-positions.
 11. The modular composition of claim 3, wherein the siRNA is double stranded; and wherein the peptides are attached to the guide strand or the passenger strand of the siRNA at different 2′-positions of the ribose rings of the siRNA.
 12. The modular composition of claim 3, wherein the siRNA is double stranded; and wherein the peptides are attached to the guide strand or the passenger strand of the siRNA at different terminal 3′ and/or 5′-positions.
 13. The modular composition of claim 3, wherein the siRNA is double stranded; and wherein the peptides are attached to both the guide strand and the passenger strand of the siRNA at different 2′-positions of the ribose rings and/or different terminal 3′ and/or 5′-positions.
 14. The modular composition of claim 3, wherein the tetraGalNAc ligands and the peptides are attached to the same strand of the siRNA.
 15. The modular composition of claim 3, wherein the tetraGalNAc ligands and the peptides are attached to different strands of the siRNA.
 16. The modular composition of claim 3, wherein the tetraGalNAc ligands and the peptides are attached to the same or different strands of the siRNA via linkers.
 17. The modular composition of claim 16, wherein each linker is independently selected from the group consisting of:

wherein: each R is independently H, Boc (tert-butyloxycarbonyl), Cbz (carboxybenzyl), Ac (acetyl), a PEG, a lipid, a targeting ligand, linker(s), or peptide(s); and each n is 0 to
 750. 18. The modular composition of claim 16, wherein each linker is independently selected from the group consisting of:

wherein: each R is independently H, Boc (tert-butyloxycarbonyl), Cbz (carboxybenzyl), Ac (acetyl), a PEG, a lipid, a targeting ligand, linker(s), or peptide(s); and each n is 0 to
 750. 19. The modular composition of claim 18, wherein the linker is a branching linker independently selected from the group consisting of:

wherein: each R is independently H, Boc (tert-butyloxycarbonyl), Cbz (carboxybenzyl), Ac (acetyl), a PEG, a lipid, a targeting ligand, linker(s), or peptide(s); and each n is 0 to
 750. 20. The modular composition of claim 3, wherein the siRNA is double stranded; and wherein the optional targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents are attached to the same or different strands of the siRNA via linkers. 